Antimicrobial 4-oxoquinolizines

ABSTRACT

This invention provides novel 4-oxoquinolizine compounds and their uses for a series of broad-spectrum antibiotics having no cross-resistance to existing or emerging classes of antibiotics. In addition the novel 4-oxoquinolizine compounds are useful against CDC Category A and B pathogens The invention also provides pharmaceutical compositions comprising certain 4-oxoquinolizines in combination with subinhibitory concentrations of polymyxin B against clinical isolates which are resistant to quinolones, carbapenems and other antimicrobial agents.

BACKGROUND OF THE INVENTION

Field of the Invention

This invention encompasses novel 2-pyridone compounds and theirpharmaceutical compositions. In particular the invention relates tonovel 4-oxoquinolizine compounds and their pharmaceutical compositions.In certain embodiments, the invention is directed to 4-oxoquinolizinesin combination with subinhibitory concentrations of polymyxin B.

Description of Related Art

Second generation quinolones such as Ciprofloxacin are widely acceptedfor the treatment of bacterial infections of the respiratory and urinarytracts, skin and soft tissues. They have good pharmacokinetic profiles,potent activities against a wide range of Gram-positive andGram-negative pathogens, and are widely used in both hospital andcommunity settings. However, increasing frequency of bacterialresistance to quinolones has led to an urgent need for new analogs toovercome antibiotic resistance.

2-pyridones have the potential to exhibit a new mechanism of action withbroad-spectrum antibacterial activity and favorable drug-like propertiesto become the first 2-pyridone members in such clinical use. 2-pyridonesare distantly related to quinolones, but with a different heterocyclicnucleus and different electronic distribution over the molecule leadingto significant changes in chemical reactivity.

In 1994, Abbott reported that 2-pyridone analogs were efficaciousagainst certain quinolone resistant microorganisms (34^(th) InterscienceConference on Antimicrobial Agents and Chemotherapy (ICAAC, paper F41),1994; U.S. Pat. No. 5,789,591), specifically ABT-719, a4-oxo-quinolizine (also described herein as compound 10)

possessing potent antibacterial activity against both Gram-positive andGram-negative pathogens. More recently, Sato disclosed a family of4-oxo-quinolizines on a 2-pyridone scaffold (U.S. Pat. No. 6,525,066;U.S. Pat. No. 7,223,773) exhibiting strong antibacterial activityagainst Gram-positive and Gram-negative and anaerobic bacteria.

There is a continuing need for antimicrobial compounds that have potentactivity against many pathogens particularly multiresistant ones.

SUMMARY OF THE INVENTION

Provided herein are 2-pyridone compounds and their uses as antimicrobialagents. Preferably the 2-pyridone compounds of the invention are4-oxoquinolizine compounds.

In one aspect the invention provides pharmaceutical compositionscomprising a Polymyxin and a 4-oxoquinolizine compound represented byformula (II)

-   -   wherein    -   R₁ is hydrogen, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ haloalkyl,        —OR^(X), —N(R^(X))₂, —C(O)R^(X), —C(O)OR^(X), or —C(O)N(R^(X))₂,        wherein each Rx is independently hydrogen, C₁₋₈ alkyl, or        C₁₋₈haloalkyl; and    -   R₇ is hydrogen, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ alkoxyl, C₁₋₈        haloalkyl, heterocyclyl, —OR¹¹, —N(R¹¹)₂, or —C(O)N(R¹¹)₂,        wherein each R¹¹ is independently hydrogen, C₁₋₈ alkyl, or C₁₋₈        haloalkyl; and    -   R₈ is a C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, or        heteroaryl, wherein the cycloalkyl and heteroaryl are optionally        substituted with one to five groups that are each independently        halogen, C₁₋₈ alkyl, —OR²¹, —N(R²¹)₂, or —C(O)OR²¹, wherein each        R²¹ is independently hydrogen or C₁₋₈ alkyl; and    -   R₉ is (CH₂)_(n)—COOH or (CH₂)_(n)—COO—R₁₀, wherein n is a        integer in the range of 0 to 3 and R₁₀ is hydrogen or a carboxyl        protecting group; and    -   Y is heterocyclyl, aryl, or heteroaryl, each optionally        substituted by one to five groups that are each independently        halogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl,        heterocyclyl, aryl, heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl,        heterocyclyl(C₁₋₈)alkyl, aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl,        —R^(Y), or —C₁₋₈ alkyl-R^(Y), wherein R^(Y) is nitro, cyano,        —OR^(Y1), —SR^(Y1), —N(R^(Y1))₂, —C(O) R^(Y1), —C(O)OR^(Y1),        —C(O)N(R^(Y1))₂, —OC(O)R^(Y1), —OC(O)OR^(Y1), —OC(O)N(R^(Y1))₂,        —N(R^(Y1))C(O)R^(Y1), —N(R^(Y1))C(O)OR^(Y1),        —N(R^(Y1))C(O)N(R^(Y1))₂, —S(O)₂R^(Y1), —S(O)₂OR^(Y1),        —S(O)₂N(R^(Y1))₂, —OS(O)₂R^(Y1), —OS(O)₂OR^(Y1),        —OS(O)₂N(R^(Y1))₂, —N(R^(Y1))S(O)₂R^(Y1), —N(R^(Y1))        S(O)₂OR^(Y1), or —N(R^(Y1))S(O)₂N(R^(Y1))₂, wherein each R^(Y1)        is independently hydrogen, C₁₋₈ alkyl, or C₁₋₈ haloalkyl.

Said polymyxin may be any of the polymyxins described herein below inthe section “Polymyxin” and said 4-oxoquinolizine compound may be any ofthe 4-oxoquinolizine compounds described herein below in the sections“4-oxoquinolizines” and “Particular 4-oxoquinolizines”.

The invention also provides said pharmaceutical compositions for use inthe treatment of a bacterial infection, which for example may be any ofthe bacterial infections described herein below in the section“Bacterial infection”.

In a further aspect, the invention provides 2-pyridone compoundsrepresented by formula (I), or a pharmaceutically acceptable saltthereof:

wherein R₁ and R₂ are independently hydrogen or fluorine, R₃ and R₅ areindependently hydrogen, fluorine or chlorine, R₄ is —NH₂ or —CH₂NH₂, R₆is H or F, and R₇ is H, CF₃, CONH₂, CH₃, OCH₃, or —CN.

In certain embodiments, 2-pyridone compounds are provided having thestructure of one of compounds 1, 2, 3, 4, 5, 6 and 17, as shown herein.

In another aspect of the invention antimicrobial agents andpharmaceutical compositions thereof comprising a 2-pyridone compound areprovided.

In another aspect, the invention provides for pharmaceuticalcompositions comprising a 2-pyridone compound in combination withpolymyxin B, wherein the polymyxin B is present in a subinhibitoryconcentration.

In another aspect, the invention provides for the use of pharmaceuticalcompositions comprising the instant 2-pyridone compounds asantimicrobials.

Uses of the pharmaceutical compositions are provided herein against oneof: Burkholderia pseudomallei, Bacillus anthracis, Yersinia pestis,Francisella tularensis, and Brucella abortus, Klebsiella, Pseudomonas,Acinetobacter, Staphylococcus aureus MRSA, S. epidermidis, Streptococcusaureus, Streptococcus pneumonia, Enterococcus faecalis, Enterococcusfaecium, B. pseudomallei, Pseudomonas aeruginosa, Burkholderiathailandensi, Acinetobacter baumannii, or Acinetobacter, Escherichiacoli, and Klebsiella.

In a further aspect the invention provides kit-of-parts comprising apolymyxin, which may be any of the polymyxins described herein below inthe section “Polymyxin” and a 4-oxoquinolizine compound, which may beany of the 4-oxoquinolizine compounds described herein below in thesections “4-oxoquinolizines” and “Particular 4-oxoquinolizines”.

In another aspect the invention provides 4-oxoquinolizine compoundsrepresented by formula (I), or a pharmaceutically acceptable saltthereof:

-   -   wherein    -   R1 is hydrogen or fluorine; and    -   R3 is fluorine, —(CH₂)_(n)—NH₂ or C₁₋₃-alkyl, wherein n is an        integer in the range of 0 to 2; and    -   R4 is —(CH₂)_(n)—NH₂, —NH—(CH2)_(n)—CH₃ or C₁₋₃-alkyl, wherein n        is an integer in the range of 0 to 2; and    -   R5 is hydrogen or C₁₋₃ alkyl; and    -   R2 and R6 are hydrogen; and    -   R7 is C₁₋₃ alkyl or C₁₋₃ alkoxy    -   with the proviso that when R3 is fluorine and R4 is amine, then        R1 is fluorine.

In a still further aspect the invention provides 4-oxoquinolizinecompounds of formula (I) or a pharmaceutically acceptable salt thereof,wherein

-   -   R1 is as defined herein below in relation to formula III; and    -   R2, R3, R4, R5 and R6 each independently are hydrogen, hydroxyl,        —(CH₂)_(n)—NH—(C═O)—(CH₂)_(m)—CH₃, —(C═O)—C₁₋₈ alkyl,        —(C═O)—C₁₋₈ haloalkyl, halogen, —(CH₂)_(n)—NH₂,        —NH—(CH₂)_(n)—CH₃, C₁₋₈-alkyl or C₁₋₈ alkoxy, wherein n and m        each independently is an integer in the range of 0 to 3 and        wherein at least one of R2, R3, R4, R5 or R6 is hydroxyl,        —(CH₂)_(n)—NH—(C═O)—(CH₂)_(n)—CH₃ or —(C═O)—C₁₋₈ haloalkyl; and    -   R7 is as defined herein below in relation to formula Ill.

In yet another aspect the invention provides 4-oxoquinolizine compoundsof formula (V) or a pharmaceutically acceptable salt thereof,

-   -   wherein R₁ is hydrogen or fluorine; and    -   R₂, R₃, R₄ and R₅ each individually are selected from the group        consisting of hydrogen, (CH₂)_(n)-hydroxyl, fluorine, C₁₋₃        alkyl, —(CH₂)_(n)—NH₂, —NH—(CH₂)_(n)—CH₃ and a 5 to 6 membered        heterocyclic ring, wherein n is an integer in the range of 0 to        2; and    -   R₆ is hydrogen and    -   R₇ is C₁₋₃ alkyl or C₁₋₃ alkoxy; and    -   Q₁, Q₂ and Q₃ each individually are C or N, wherein at least one        of Q₁, Q₂ and Q₃ is N and at least one of Q₁, Q₂ and Q₃ is C,        and wherein if Q₁ is N, then R₃ is not present, and if Q₂ is N,        then R₄ is not present and if Q₃ is N, then R₅ is not present    -   with the provisos that    -   if Q₁ is N and Q₂ and Q₃ are C, then at least one of R₂, R₄ and        R₅ is not hydrogen; and    -   if Q₁ is N and Q₂ and Q₃ are C and R₄ is —NH₂, then R₁ is        fluorine and/or R₇ is methoxy; and    -   if Q₃ is N and Q₂ and Q₁ are C, then at least one of R₂, R₃ and        R₄ is not hydrogen; and    -   if Q₃ is N and Q₂ and Q₁ are C and R₄ is —NH₂, then R₁ is        fluorine and/or R₇ is methoxy.

In another aspect the invention provides 4-oxoquinolizine compounds offormula (IIIa) or pharmaceutically acceptable salts thereof, wherein

-   -   R₁ is as defined herein below in relation to compounds of        formula III; and    -   Y is a heterobicyclic ring system optionally substituted with        one or more substituents selected from the group consisting of        oxo, —(CH₂)_(n)—NH₂, —NH—(CH₂)_(n)—CH₃, —(CH₂)_(n)—OH, C₁₋₈        alkyl, C₁₋₈ alkoxy, C₃₋₈-cycloalkyl and halogen, wherein n is an        integer in the range of 0 to 3; and    -   R₇ is as defined herein below in relation to compounds of        formula III.

In an even further aspect the invention relates to 4-oxoquinolizinecompounds of formula (IIIa) or pharmaceutically acceptable saltsthereof, wherein

-   -   R₁ is as defined herein below in relation to compounds of        formula III; and    -   Y is selected from the group consisting of pyrazolyl and        tetrahydropyrinidyl optionally substituted with one or more        substituents selected from the group consisting of oxo,        —(CH₂)_(n)—NH₂, —NH—(CH₂)_(n)—CH₃, —(CH₂)_(n)—OH, C₁₋₈ alkyl,        C₁₋₈ alkoxy, C₃₋₈-cycloalkyl and halogen, wherein n is an        integer in the range of 0 to 3; and    -   R₇ is as defined herein below in relation to compounds of        formula III.

These and other features and advantages of the present invention will bemore fully understood from the following detailed description of theinvention taken together with the accompanying claims. It is noted thatthe scope of the claims is defined by the recitations therein and not bythe specific discussion of features and advantages set forth in thepresent description.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of the embodiments of the presentinvention can be best understood when read in conjunction with thefollowing drawings.

FIG. 1 shows the Isobologram related to the synergy of the antibacterialactivity of compounds 2, 33 and 35 with Polymyxin B on the AcinetobacterGN52 strain.

FIG. 2 shows the Isobologram related to the synergy of the antibacterialactivity of compounds 2, 33 and 35 with Polymyxin B on the AcinetobacterGN56 strain.

FIG. 3 shows the Isobologram related to the synergy of the antibacterialactivity of compounds 2 and 35 with Polymyxin B on the Klebsiellapneumonia-NDM-1 strain.

FIGS. 4A-4E show the structures of the 5 scaffolds used as intermediatestowards the preparation of the 4-oxoquinolizines compounds

FIG. 5A is an LC-MS characterization of compound 1. The data wasrecorded on a Waters Acquity UPLC system, equipped with SQD, PDA andELSD detectors, with an Acquity BEH C18 1.7 micron column.

FIG. 5b is an LC characterization of compound 1. The LC was run with a10 min gradient from 0 to 100% B (A: water with 0.1% formic acid, B:acetonitrile with 0.1% formic acid).

FIG. 6A is an LC-MS characterization of compound 5. The data wasrecorded on a Waters Acquity UPLC system, equipped with SQD, PDA andELSD detectors, with an Acquity BEH C18 1.7 micron column.

FIG. 6B is an LC characterization of compound 5. The LC was run with a10 min gradient from 0 to 100% B (A: water with 0.1% formic acid, B:acetonitrile with 0.1% formic acid).

FIG. 7A is an LC-MS characterization of compound 6. The data wasrecorded on a Waters Acquity UPLC system, equipped with SQD, PDA andELSD detectors, with an Acquity BEH C18 1.7 micron column.

FIG. 7B is an LC characterization of compound 6. The LC was run with a10 min gradient from 0 to 100% B (A: water with 0.1% formic acid, B:acetonitrile with 0.1% formic acid).

Elements in the figures are illustrated for simplicity and clarity andhave not necessarily been drawn to scale. For example, the dimensions ofsome of the elements in the figures can be exaggerated relative to otherelements to help improve understanding of the embodiment(s) of thepresent invention.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Specific compounds are named herein as Compound followed by a number,i.e. Compound n, wherein n is an integer. This refers to the compoundsas named and numbered in Table 1 herein below.

The term “alkyl” as used herein refers to a saturated, straight orbranched hydrocarbon chain. The hydrocarbon chain preferably contains offrom one to eight carbon atoms (C₁₋₈ alkyl), such preferably from one tosix carbon atoms (C₁₋₆ alkyl), more preferred of from one to five carbonatoms (C₁₋₅-alkyl), including methyl, ethyl, propyl, isopropyl, butyl,isobutyl, secondary butyl, tertiary butyl, pentyl, isopentyl, neopentyland tertiary pentyl. In a preferred embodiment alkyl represents aC₁₋₄-alkyl group, which may in particular include methyl, ethyl, propyl,isopropyl, butyl, isobutyl, secondary butyl, and tertiary butyl.

The term “aryl” as used herein refers to an aromatic ring or aromaticring system substituent. Aryl may for example be phenyl or naphthyl.

The term “cycloalkyl” as used herein refers to a cyclic alkyl group,preferably containing of from three to eight carbon atoms(C₃₋₈-cycloalkyl), including cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl and cyclooctyl, more preferably 3 carbon atoms(cyclopropyl).

The term “haloalkyl” as used herein refers to an alkyl group as definedherein, which alkyl group is substituted one or more times with one ormore halogen.

The term “heteroaryl” refers to an aryl, wherein one or more ringcarbons have been exchanged for a heteroatom. The heteroatom is ingeneral selected from the group consisting of N, S and O. The heteroarylpreferably contains 1 to 3 heteroatoms.

The term “heterocyclyl” as used herein refers to a monocyclic group or amulticyclic group holding one or more heteroatoms in its ring structure.Preferably heterocyclyl refers to monocyclic or bicyclic groups.Preferred heteroatoms include nitrogen (N), oxygen (O) and sulphur (S).Examples of 5-membered monocyclic heterocyclic groups includepyrrolidinyl, pyrrolyl, 3H-pyrrolyl, oxolanyl, furanyl, thiolanyl,thiophenyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolidinyl,1,2-oxazolyl, 1,3-oxazolyl, 1,2-thiazolyl, 1,3-thiazolyl, and1,2,5-oxadiazolyl. Examples of 6-membered monocyclic heterocyclic groupsinclude piperidinyl, pyridinyl, oxanyl, 2-H-pyranyl, 4-H-pyranyl,thianyl, 2H-thiopyranyl, pyridazinyl, 1,2-diazinanyl, pyrimidinyl,1,3-diazinanyl, pyrazinyl, piperazinyl, 1,4-dioxinyl, 1,4-dioxanyl,1,4-oxazinyl, morpholinyl, thiomorpholinyl and 1,4-oxathianyl.

The term “bicyclic heteroaryl” as used herein refers to a bicyclicaromatic ring system substituent derived by fusion of two monocyclicgroups, where at least one of said two monocyclic groups holds one ormore heteroatoms in its ring structure. Preferred heteroatoms includenitrogen (N), oxygen (O) and sulphur (S). Examples of bicyclicheterocyclic group includes 1H-indolyl, decahydroquinolinyl,octahydrocyclopenta[b]pyrrolyl, 4H-chromenyl,2,3-dihydro-1-benzofuranyl, 2H-1,3-benzodioxolyl, 1H-1,3-benzodiazolyland 1,3-benzothiazolyl.

The term “halogen” as used herein refers to a substituent selected fromthe group consisting of —Cl, —F, —Br and —I.

The term “oxo” as used herein refers to ═O.

4-Oxoquinolizine Compounds

The instant invention provides novel 2-pyridones having potency, breadthof antimicrobial activity, lack of cross-resistance to existing drugs,safety, and/or efficacy in animal models for Category A & B CDCpathogens including the particularly problematic bacteria residinginside mammalian host cells. The preferred 2-pyridone compounds of theinvention are 4-oxoquinolizine compounds. The disclosed 2-pyridonecompounds have substituted 2-pyridone scaffolds and preferably the2-pyridone compounds contain a substituted 4-oxoquinolizine scaffold.They target the well validated IIA bacterial topoisomerases, e.g. DNAgyrase and topoisomerase IV, thereby inhibiting a broad spectrum ofGram-positive as well as Gram-negative bacteria—importantly, thoseresistant to quinolones and topoisomerase inhibitors, such as forexample piperidinylalkylquinolines. By synthesis and profiling in vitroand in vivo, the presently disclosed series of 4-oxoquinolizineantibiotics have been found to be broad-spectrum and very potent againstCDC pathogens, including Burkholderia pseudomallei, Bacillus anthracis,Yersinia pestis, Francisella tularensis, and Brucella abortus in MIC90ranges of low nanograms per milliliter. Similar MICs are obtainedagainst important nosocomial pathogens, among them quinolone-resistantKlebsiella, Pseudomonas, Acinetobacter and Staphylococcus aureus MRSA.Furthermore, we have found that 4-oxoquinolizines are equally effectiveagainst a surrogate Burkholderia strain replicating inside mammalianhost cells.

2-pyridones are distantly related to quinolones, but with a differentheterocyclic nucleus and different electronic distribution over themolecule leading to significant changes in chemical reactivity.Exhibiting a possibly new mechanism of action against a well-validatedbacterial target, with favorable drug-like properties, the instant2-pyridones are likely to become the first 2-pyridone members in suchclinical use. In particular, the 2-pyridones may be 4-oxoquinolizinecompounds.

It is one aspect of the present invention to provide pharmaceuticalcompositions comprising 2-pyridone compounds in combination withpolymyxin. Said 2-pyridone compound is preferably a 4-oxoquinolizinecompound and said Polymyxin is preferably present in a subinhibitoryconcentration and is preferably Polymyxin B as described hereinelsewhere.

The 2-pyridone compound according to the present invention may be anycompound having a 2-pyridone skeleton as a partial structure. In apreferred embodiment the 2-pyridone compound is a 4-oxoquinolizinecompound, i.e. a compound comprising a 4-oxoquinolizine skeletonstructure.

Thus, in a preferred embodiment of the invention the present inventionrelates to pharmaceutical compositions comprising a 4-oxoquinolizinecompound and polymyxin, wherein said polymyxin preferably is present insubinhibitory concentrations.

The 4-oxoquinolizine compound is preferably a compound of the formula(II):

or a pharmaceutically acceptable salt thereof, wherein

-   R₁ is hydrogen, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ haloalkyl, —OR^(X),    —N(R^(X))₂, —C(O)R^(X), —C(O)OR^(X), or —C(O)N(R^(X))₂, wherein each    Rx is independently hydrogen, C₁₋₈ alkyl, or C₁₋₈haloalkyl; and-   R₇ is hydrogen, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ alkoxyl, C₁₋₈    haloalkyl, heterocyclyl, —OR¹¹, —N(R¹¹)₂, or —C(O)N(R¹¹)₂, wherein    each R¹¹ is independently hydrogen, C₁₋₈ alkyl, or C₁₋₈ haloalkyl;    and-   R₈ is a C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, or heteroaryl,    wherein the cycloalkyl and heteroaryl are optionally substituted    with one to five groups that are each independently halogen, C₁₋₈    alkyl, —OR²¹, —N(R²¹)₂, or —C(O)OR²¹, wherein each R²¹ is    independently hydrogen or C₁₋₈ alkyl; and-   R₉ is (CH₂)_(n)—COOH or (CH₂)_(n)—COO—R₁₀, wherein n is a integer in    the range of 0 to 3 and R₁₀ is hydrogen or a carboxyl protecting    group; and-   Y is heterocyclyl, aryl, or heteroaryl, each optionally substituted    by one to five groups that are each independently halogen, C₁₋₈    alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,    heteroaryl, C₃₋₈ cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,    aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈    alkyl-R^(Y), wherein R^(Y) is nitro, cyano, —OR^(Y1), —SR^(Y1),    —N(R^(Y1))₂, —C(O)R^(Y1), —C(O)OR^(Y1), —C(O)N(R^(Y1))₂,    —OC(O)R^(Y1), —OC(O)OR^(Y1), —OC(O)N(R^(Y1))₂, —N(R^(Y1))C(O)R^(Y1),    —N(R^(Y1))C(O)OR^(Y1), —N(R^(Y1))C(O)N(R^(Y1))₂, —S(O)₂R^(Y1),    —S(O)₂OR^(Y1), —S(O)₂N(R^(Y1))₂, —OS(O)₂R^(Y1), —OS(O)₂OR^(Y1),    —OS(O)₂N(R^(Y1))₂, —N(R^(Y1))S(O)₂R^(Y1), —N(R^(Y1))S(O)₂OR^(Y1), or    —N(R^(Y1))S(O)₂N(R^(Y1))₂, wherein each R^(Y1) is independently    hydrogen, C₁₋₈ alkyl, or C₁₋₈ haloalkyl.

In relation to compounds of formula (II) it is preferred that R₉ is(CH₂)_(n)—COOH, and more preferably R₉ is —COOH.

More preferably the 4-oxoquinolizine compound is a compound of formula(III):

or a pharmaceutically acceptable salt thereof, wherein

-   R₁ is hydrogen, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ haloalkyl, —OR^(X),    —N(R^(X))₂, —C(O)R^(X), —C(O)OR^(X), or —C(O)N(R^(X))₂, wherein each    Rx is independently hydrogen, C₁₋₈ alkyl, or C₁₋₈haloalkyl; and-   R₇ is hydrogen, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ alkoxyl, C₁₋₈    haloalkyl, heterocyclyl, —OR¹¹, —N(R¹¹)₂, or —C(O)N(R¹¹)₂, wherein    each R¹¹ is independently hydrogen, C₁₋₈ alkyl, or C₁₋₈ haloalkyl;-   R₈ is a C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, or heteroaryl,    wherein the cycloalkyl and heteroaryl are optionally substituted    with one to five groups that are each independently halogen, C₁₋₈    alkyl, —OR²¹, —N(R²¹)₂, or —C(O)OR²¹, wherein each R²¹ is    independently hydrogen or C₁₋₈ alkyl;-   Y is heterocyclyl, aryl, or heteroaryl, each optionally substituted    by one to five groups that are each independently halogen, C₁₋₈    alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,    heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,    aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈    alkyl-R^(Y), wherein R^(Y) is nitro, cyano, —OR^(Y1), —SR^(Y1),    —N(R^(Y1))₂, —C(O)R^(Y1), —C(O)OR^(Y1), —C(O)N(R^(Y1))₂,    —OC(O)R^(Y1), —OC(O)OR^(Y1), —OC(O)N(R^(Y1))₂, —N(R^(Y1))C(O)R^(Y1),    —N(R^(Y1))C(O)OR^(Y1), —N(R^(Y1))C(O)N(R^(Y1))₂, —S(O)₂R^(Y1),    —S(O)₂OR^(Y1), —S(O)₂N(R^(Y1))₂, —OS(O)₂R^(Y1), —OS(O)₂OR^(Y1),    —OS(O)₂N(R^(Y1))₂, —N(R^(Y1))S(O)₂R^(Y1), —N(R^(Y1))S(O)₂OR^(Y1), or    —N(R^(Y1))S(O)₂N(R^(Y1))₂, wherein each R^(Y1) is independently    hydrogen, C₁₋₈ alkyl, or C₁₋₈ haloalkyl.

In one embodiment of the invention the 4-oxoquinolizine compound is acompound of formula IIIa:

or a pharmaceutically acceptable salt thereof, wherein the R₁, R₇ and Yare as defined herein above in relation to compounds of formula (III).

In another embodiment of the invention the 4-oxoquinolizine compound isa compound of formula IIIIb:

or a pharmaceutically acceptable salt thereof, wherein the R₁, R₈ and Yare as defined herein above in relation to compounds of formula (III).

In another embodiment of the invention the 4-oxoquinolizine compound isa compound of formula IIIIc:

or a pharmaceutically acceptable salt thereof, wherein the R₁ and Y areas defined herein above in relation to compounds of formula (III).

In relation to compounds according to formula (II). (III) and (IIIa) theR₇ may preferably be as described in the following paragraph:

In one preferred embodiment R₇ is hydrogen, halogen, or C₁₋₈ alkyl, andmore preferably R₇ is hydrogen or C₁₋₈ alkyl, yet more preferably R₇ ishydrogen or C₁₋₅ alkyl, yet more preferably R₇ is hydrogen or C₁₋₂alkyl, yet more preferably R₇ is hydrogen or methyl. In anotherembodiment, R₇ is halogen or C₁₋₈ alkyl. In another embodiment, R₇ isC₁₋₈ alkyl, preferably R₇ is C₁₋₅ alkyl, yet more preferably R₇ is C₁₋₂alkyl, yet more preferably R₇ is methyl. In another embodiment, R₇ ishalogen. In another embodiment R₇ is C₁₋₈ haloalkyl, —OR¹¹, or—C(O)N(R¹)₂. In another embodiment R₇ is trifluoromethyl, methoxy, or—C(O)NH₂.

In relation to compounds of formula (II), (III), (IIIa), (IIIb) and(IIIc), then R₇ may be as described herein above and R₁ may preferablybe as described in the following paragraph: In one preferred embodimentR₁ may be hydrogen or halogen. Thus in one very preferred embodiment R₁is hydrogen. In another preferred embodiment R₁ is halogen, and morepreferably R₁ may be fluorine. In another embodiment, R₁ is hydrogen,halogen, C₁₋₈ alkyl, C₁₋₈ alkoxy, amino, C₁₋₈ alkylamino, or di(C₁₋₈alkyl)amino, preferably R₁ may be C₁₋₈ alkyl, more preferably C₁₋₅alkyl, even more preferably C₁₋₂ alkyl, yet more preferably methyl. Inanother embodiment, R₁ is —OR^(X), —N(R^(X))₂, —C(O)R^(X), —C(O)OR^(X),or —C(O)N(R^(X))₂. In another embodiment, R₁ is —OR^(X) or —N(R^(X))₂.In another embodiment, R₁ is —C(O)R^(X), —C(O)OR^(X), or —C(O)N(R^(X))₂.In another embodiment, R₁ is C₁₋₈haloalkyl (e.g., trifluoromethyl).

In particular preferred embodiments of the invention the compounds offormula (II), (III) and (IIIa) have R₇ and R₁ selected from one of thefollowing combinations:

-   -   (a) R₇ is hydrogen, halogen, or C₁₋₈ alkyl; and R₁ is hydrogen        or halogen.    -   (b) R₇ is hydrogen, halogen, or C₁₋₈ alkyl; and R₁ is hydrogen.    -   (c) R₇ is hydrogen, halogen, or C₁₋₈ alkyl; and R₁ is halogen.    -   (d) R₇R¹ is hydrogen, halogen, or C₁₋₈ alkyl; and R₁ is fluoro.    -   (e) R₇ is hydrogen or C₁₋₈ alkyl; and R₁ is hydrogen or halogen.    -   (f) R₇ is hydrogen or C₁₋₈ alkyl; and R₁ is hydrogen.    -   (g) R₇ is hydrogen or C₁₋₈ alkyl; and R₁ is halogen.    -   (h) R₇ is hydrogen or C₁₋₈ alkyl; and R₁ is fluoro.    -   (i) R₇ is hydrogen or methyl; and R₁ is hydrogen or halogen.    -   (j) R₇ is hydrogen or methyl; and R₁ is hydrogen.    -   (k) R₇ is hydrogen or methyl; and R₁ is halogen.    -   (l) R₇ is hydrogen or methyl; and R₁ is fluoro.    -   (m) R₇ is halogen or C₁₋₈ alkyl; and R₁ is hydrogen or halogen.    -   (n) R₇ is halogen or C₁₋₈ alkyl; and R₁ is hydrogen.    -   (o) R₇ is halogen or C₁₋₈ alkyl; and R₁ is halogen.    -   (p) R₇ is halogen or C₁₋₈ alkyl; and R₁ is fluoro.    -   (q) R₇ is C₁₋₈ alkyl; and R₁ is hydrogen or halogen.    -   (r) R₇ is C₁₋₈ alkyl; and R₁ is hydrogen.    -   (s) R₇ is C₁₋₈ alkyl; and R₁ is halogen.    -   (t) R₇ is C₁₋₈ alkyl; and R₁ is fluoro.

In relation to compounds of formula (II), (III), (IIIa), (IIIb) and(IIIc), then R₁ and R₇ may preferably be as described herein above and Ymay preferably be as described in the following paragraphs 1) to 19):

-   -   1) In one embodiment Y is aryl optionally substituted by one to        five groups that are each independently halogen, C₁₋₈ alkyl,        C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl, heteroaryl,        C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,        aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), C(O)R^(Y1), or        —C₁₋₈ alkyl-R^(Y).    -   2) In another embodiment Y is phenyl substituted with one group        which is halogen, cyano, —OR^(Y1), —SR^(Y1), —N(R^(Y1))₂, C₁₋₈        alkyl, —C₁₋₈ alkyl-N(R^(Y1))₂, or —C₁₋₈ alkyl-OR^(Y1), and        optionally substituted by one to two groups that are each        independently halogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl,        C₃₋₈cycloalkyl, heterocyclyl, aryl, heteroaryl,        C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,        aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈        alkyl-R^(Y).    -   3) In another embodiment Y is phenyl substituted with one group        which is cyano, —OR^(Y1), —N(R^(Y1))₂, C₁₋₈ alkyl, or —C₁₋₈        alkyl-N(R^(Y1))₂, and optionally substituted by one to two        groups that are each independently halogen, C₁₋₈ alkyl, —R^(Y),        or —C₁₋₈ alkyl-R^(Y).    -   4) In another embodiment Y is phenyl substituted with one group        which is —N(R^(Y1))₂ or —C₁₋₈ alkyl-N(R^(Y1))₂, and optionally        substituted by one to two groups that are each independently        halogen, C₁₋₈ alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y).    -   5) In another embodiment Y is phenyl substituted with one group        which is —NH₂ or —C₁₋₈ alkyl-NH₂, and optionally substituted by        one to two groups that are each independently halogen, C₁₋₈        alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y).    -   6) In another embodiment Y is phenyl substituted with one group        which is —NH₂ or —CH₂NH₂, and optionally substituted by one to        two groups that are each independently halogen, C₁₋₈ alkyl,        —R^(Y), or —C₁₋₈ alkyl-R^(Y).    -   7) In another embodiment Y is phenyl substituted with one group        which is —NH₂ or —CH₂NH₂, and substituted by one to two groups        that are each independently halogen, C₁₋₈ alkyl, —R^(Y), or        —C₁₋₈ alkyl-R^(Y).    -   8) In another embodiment Y is heteroaryl optionally substituted        by one to five groups that are each independently halogen, C₁₋₈        alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,        heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,        aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈        alkyl-R^(Y).    -   9) In another embodiment Y is a 5-membered or 6-membered        heteroaryl optionally substituted by one to five groups that are        each independently halogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl,        C₃₋₈cycloalkyl, heterocyclyl, aryl, heteroaryl,        C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,        aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈        alkyl-R^(Y).    -   10) In another embodiment Y is a 5-membered or 6-membered        heteroaryl substituted with one group which is —N(R^(Y1))₂ or        —C₁₋₈ alkyl-N(R^(Y1))₂, and optionally substituted by one to two        groups that are each independently halogen, C₁₋₈ alkyl, —R^(Y),        or —C₁₋₈ alkyl-R^(Y).    -   11) In another embodiment Y is pyrazolyl,    -   12) In another embodiment Y is pyridyl, furyl, or thienyl each        optionally substituted with one group which is —N(R^(Y1))₂ or        —C₁₋₈ alkyl-N(R^(Y1))₂, and each optionally substituted by one        to two groups that are each independently halogen, C₁₋₈ alkyl,        —R^(Y), or —C₁₋₈ alkyl-R^(Y).    -   13) In another embodiment Y is pyridyl, furyl, or thienyl each        optionally substituted with one group which is —NH₂ or —CH₂NH₂,        and each optionally substituted by one to two groups that are        each independently halogen, C₁₋₈ alkyl, —R^(Y), or —C₁₋₈        alkyl-R^(Y).    -   14) In another embodiment Y is pyridyl, tetrahydro-pyridinyl or        pyrimidinyl optionally substituted with one group, which is        halogen, C₁₋₈ alkyl or R^(Y).    -   15) In another embodiment Y is a bicyclic heteroaryl optionally        substituted by one to five groups that are each independently        halogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl,        heterocyclyl, aryl, heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl,        heterocyclyl(C₁₋₈)alkyl, aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl,        —R^(Y), or —C₁₋₈ alkyl-R^(Y).    -   16) In another embodiment Y is a benzofuranyl, benzothienyl,        indolyl, indazolyl, benzimidazolyl, benzoxazolyl,        benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, or        benzotriazolyl, each optionally substituted by one to five        groups that are each independently halogen, C₁₋₈ alkyl, C₁₋₈        haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl, heteroaryl,        C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,        aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈        alkyl-R^(Y).    -   17) In another embodiment Y is indazolyl optionally substituted        by one to five groups that are each independently halogen, C₁₋₈        alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,        heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,        aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈        alkyl-R^(Y).    -   18) In another embodiment Y is indazolyl.    -   19) In another embodiment Y is indolinyl or pyrrolopyridinyl        optionally substituted with oxo.

In one preferred embodiment of the invention, then Y in relation tocompounds of formula (II), (III), (IIIa), (IIIb) and (IIIc) is phenyl orpyridyl substituted with —(CH₂)_(n)—NH₂ or —(CH₂)_(n)—NH—(CH₂)_(m)—CH₃,wherein n and m independently are integers in the range of 0 to 3,wherein said phenyl or said pyridyl optionally may be substituted withone or two additional substituents selected from the group consisting ofhalogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y),wherein R^(Y) is nitro, cyano, —OR^(Y1), —SR^(Y1), —N(R^(Y1))₂,—C(O)R^(Y1), —C(O)OR^(Y1), —C(O)N(R^(Y1))₂, —OC(O)R^(Y1), —OC(O)OR^(Y1),—OC(O)N(R^(Y1))₂, —N(R^(Y1))C(O)R^(Y1), —N(R^(Y1))C(O)OR^(Y1),—N(R^(Y1))C(O)N(R^(Y1))₂, —S(O)₂R^(Y1), —S(O)₂O R^(Y1),—S(O)₂N(R^(Y1))₂, —OS(O)₂R^(Y1), —OS(O)₂OR^(Y1), —OS(O)₂N(R^(Y1))₂,—N(R^(Y1))S(O)₂R^(Y1), —N(R^(Y1))S(O)₂OR^(Y1), or—N(R^(Y1))S(O)₂N(R^(Y1))₂, wherein each R^(Y1) is independentlyhydrogen, C₁₋₈ alkyl, or C₁₋₈ haloalkyl.

In this embodiment it is preferred that Y in relation to compounds offormula (II), (Ill), (IIIa), (IIIb) and (IIIc) is phenyl substitutedwith —(CH₂)_(n)—NH₂ or —(CH₂)_(n)—NH—(CH₂)_(m)—CH₃, wherein n and mindependently are integers in the range of 0 to 1,

wherein said phenyl may optionally be substituted with one or twoadditional substituents selected from the group consisting of halogen,C₁₋₈ alkyl, C₁₋₈ haloalkyl, nitro, cyano and hydroxyl.

In relation to compounds of formula (II), (III) and (IIIb), then R₁, R₇and Y are preferably as described herein above and R₈ may preferably beas described in the following paragraph: In a preferred embodiment R₈ isa C₁₋₈ alkyl, C₁₋₈ haloalkyl, or C₃₋₈cycloalkyl. In another embodimentR₈ is a C₁₋₈ alkyl (e.g., methyl). In another embodiment R₈ is a C₁₋₈haloalkyl (e.g., trifluoromethyl). In a very preferred embodiment R₈ isC₃₋₈cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl), and more preferably cyclopropyl or cyclobutyl, yet morepreferably cyclopropyl. In one embodiment R₈ is C₃₋₈cycloalkyloptionally substituted by one or two groups that are each independentlyhalogen or C₁₋₈ alkyl (e.g., fluoro or methyl). In another embodiment R₈is heteroaryl, wherein the heteroaryl is optionally substituted with oneto five groups that are each independently halogen, C₁₋₈ alkyl, —OR²¹,—N(R²¹)₂, or —C(O)OR²¹, wherein each R²¹ is independently hydrogen orC₁₋₈ alkyl.

In one embodiment of the invention the 4-oxoquinolizine compound is acompound of the formula (IV):

or a pharmaceutically acceptable salt thereof, whereinR₁ is hydrogen or a halogen; andR₇ is hydrogen, halogen, a lower alkyl, a lower alkoxyl or a hydroxyl;andR₁₀ is hydrogen or a carboxyl protecting group; andY is a phenyl or an aromatic group selected from the group consisting of5 membered or 6-membered heterocyclic groups each optionally substitutedwith a group selected from the group consisting of lower alkyl, loweralkoxy, nitro, cyano, amino, acyl, carbamoyl, ureido, halogen, hydroxyland carboxyl.

In particular, the 4-oxoquinolizine compound may be any of the4-oxoquinolizine compounds described in US patent applicationUS2004/0229903, the content of which is hereby incorporated byreference. More particular, the 4-oxoquinolizine compounds may beselected from the group of compounds described in Examples 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122 and 123 ofUS2004/0229903.

In a preferred embodiment of the invention the 4-oxoquinolizine compoundis selected from the group of compounds mentioned in Table 1 hereinbelow.

TABLE 1 Examples of 4-oxoquinolizines compounds Compounds Structure NameCompound 1

8-(3-fluoro-4-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 2

8-(3,6-difluoro-4-amino- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 3

8-(3,5-dichloro-4-amino- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 4

8-(3-fluoro-4-aminomethyl- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 5

8-(4-aminomethyl-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 6

8-(4-aminomethyl-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 7

8-(4-amino-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 8

8-(4-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 9

8-(3-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 10

8-[(3S)-3-aminocyclopentyl]-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 11

8-(2-chloro-4-amino-5-methyl- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 12

8-[5-aminomethyl)-2-furyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 13

8-[5-aminomethyl)-2-thienyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 14

8-(4-cyanophenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 15

8-(p-tolyl)-1-cyclopropyl-9- methyl-4-oxo-quinolizine-3- carboxylic acidCompound 16

8-(4-amino-3-ethyl-5-methyl- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 17

8-(3-fluoro-4-amino-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 18

8-(3-amino-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 19

8-(4-carbamoyl-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 20

8-(2-fluoro-4-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 21

8-(3-amino-4-fluoro-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 22

8-(3-amino-5-fluoro-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 23

8-(3-fluoro-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 24

8-(3-chloro-4-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 25

8-(3-methoxy-4-amino-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 26

8-(4-acetamido-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 27

8-(4-sulfonamido-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 28

8-(4-methylamino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 29

8-(4-pyridyl)-1-cyclopropyl-9- methyl-4-oxo-quinolizine-3- carboxylicacid Compound 30

8-(3-pyridyl)-1-cyclopropyl-9- methyl-4-oxo-quinolizine-3- carboxylicacid Compound 31

8-(3-methyl-4-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 32

8-(2-fluoro-4-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 33

8-(6-amino-3-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 34

8-(1H-indol-5-yl)-1-cyclopropyl- 9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 35

8-(1H-indazol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 36

8-(4-ureido-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 37

8-(4-dimethylamino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 38

8-[(3S)-3-aminopyrrolidin-1-yl]- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 39

8-(piperazin-1-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 40

8-[(3S)-3-amino-1-piperidyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 41

8-(4-carbamoyl-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 42

8-(4-carboxy-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 43

8-(2,5-difluoro-4-amino- phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 44

8-(3,5-difluoro-4-amino- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 45

8-(3-fluoro-4-cyano-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 46

8-(4-carboxy-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 47

8-(1,2,3,6-tetrahydro-pyridin-4- yl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 48

8-(1H-pyrrol-3-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 49

8-(4-fluoro-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 50

8-(4-chloro-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 51

8-(4-hydroxy-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 52

8-(4-methoxy-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 53

8-(4-hydroxymethyl-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 54

8-(3-amino-2-oxo-3,4-dihydro- 1H-quinolin-7-yl)-1-cyclopropyl-9-methyl-4-oxo- quinolizine-3-carboxylic acid Compound 55

8-(6-hydroxy-3-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 56

8-(4-hydroxyphenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 57

8-(2-aminopyrimidin-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 58

8-(3-fluoro-4-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 59

8-(4-pyridyl)-1-cyclopropyl-7- fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 60

8-(6-amino-3-pyridyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 61

8-(4-hydroxyphenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 62

8-(1,2,3,6-tetrahydro-pyridin-4- yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 63

8-[4-(2,2,2- trifluoroacetyl)phenyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 64

8-[4- (acetamidomethyl)phenyl]--1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 65

8-(3-methyl-4-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 66

8-(3-methyl-4-hydroxyphenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 67

8-(1H-pyrazol-4-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 68

8-(3-oxoisoindolin-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 69

8-[3-methyl-4- (methylamino)phenyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 70

8-(3-fluoro-4-hydroxy-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 71

8-(3-cyclopropyl-1H-indazol-5- yl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 72

8-[3-(aminomethyl)-4-hydroxy- phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 73

8-(2-amino-1,3-benzothiazol-5- yl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 74

8-(1H-benzimidazol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 75

8-(1H-indazol-5-yl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 76

8-[3-(aminomethyl)-4-amino- phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 77

8-(indolin-5-yl)-1-cyclopropyl-9- methyl-4-oxo-quinolizine-3- carboxylicacid Compound 78

8-[6-(methylamino)-3-pyridyl]- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 79

8-(6-amino-5-methyl-3-pyridyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 80

8-(3-methyl-1H-indazol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 81

8-(1-methylindazol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 82

8-(1H-indazol-4-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 83

8-(1H-indazol-6-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 84

8-(6-piperazin-1-yl-3-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 85

8-(1H-pyrrolo-[2,3-b]-pyridin-5- yl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 86

8-(3-amino-1H-indazol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 87

8-(6-amino-3-pyridyl)-1- cyclopropyl-7,9-dimethyl-4-oxo-quinolizine-3-carboxylic acid Compound 88

8-(1H-indazol-5-yl)-7,9- dimethyl-4-oxo-quinolizine-3- carboxylic acidCompound 89

8-(6-amino-3-pyridyl)-1- cyclopropyl-9-methoxy-4-oxo-quinolizine-3-carboxylic acid Compound 90

8-(1H-indazol-5-yl)-9-methoxy- 4-oxo-quinolizine-3-carboxylic acid

Particular 4-Oxoquinolizines

It is one aspect of the present invention to provide pharmaceuticalcompositions comprising a 2-pyridone compound, preferably a4-oxoquinolizine compound in combination with a Polymyxin, preferablyPolymyxin B. Said 4-oxoquinolizine compound may be any of the4-oxoquinolizine compounds described herein above in the section4-oxoquinolizines, however it may also be any of the 4-oxoquinolizinecompounds described in this section.

It is furthermore an aspect of the present invention to provideparticularly useful 2-pyridines, i.e. such as particularly useful4-oxoquinolizine compounds. Said particularly useful 4-oxoquinolizinecompounds have a strong antibacterial effect, i.e. that they are usefulin the treatment of bacterial infections.

The particular 4-oxoquinolizine compounds are preferably a compoundhaving activity against a pathogen, having the structure of formula (I)

or a pharmaceutically acceptable salt thereof.

In one embodiment of the invention the compounds of formula (I) have Rgroups as follows: R1 and R2 are independently hydrogen or fluorine, R3and R5 are independently hydrogen, fluorine or chlorine, R4 is —NH₂ or—CH₂NH₂, R6 is H or F, and R7 is H, CF₃, CONH₂, CH₃, OCH₃, or —CN.

In another embodiment of the invention the preferred 4-oxoquinolizinecompounds are compounds of formula (I) or pharmaceutically acceptablesalts thereof, wherein

R1 is hydrogen or fluorine; andR3 is fluorine, —(CH₂)_(n)—NH₂ or C₁₋₃-alkyl, wherein n is an integer inthe range of 0 to 2; andR4 is —(CH₂)_(n)—NH₂, —NH—(CH2)_(n)—CH₃ or C₁₋₃-alkyl, wherein n is aninteger in the range of 0 to 2; andR5 is hydrogen or C₁₋₃ alkyl; andR2 and R6 are hydrogen; andR7 is C₁₋₃ alkyl or C₁₋₃ alkoxy.

In this embodiment it is preferred that

R1 is hydrogen or fluorine; andR3 is fluorine, —NH₂, —CH₂—NH₂ or methyl; andR4 is —NH₂, —(CH₂)—NH₂, —NH—CH₃ or methyl; andR5 is hydrogen or ethyl; andR2 and R6 are hydrogen; andR7 is methyl.

It is furthermore preferred that when R3 is fluorine and R4 is amine,then R1 is fluorine.

Examples of useful 4-oxoquinolizine compounds according to thisembodiment may for example be selected from the group consisting ofcompounds 4, 16, 17, 69 and 76 of Table 1.

In another embodiment of the present invention the preferred4-oxoquinolizine compounds of the invention are compounds of formula (I)or a pharmaceutically acceptable salt thereof, wherein

R1 is as defined herein above in relation to compounds of formula (III);andR2, R3, R4, R5 and R6 each independently are hydrogen, hydroxyl,—(CH₂)_(n)—NH—(C═O)—(CH₂)_(m)—CH₃, —(C═O)—C₁₋₈ alkyl, —(C═O)—C₁₋₈haloalkyl, halogen, —(CH₂)_(n)—NH₂, —NH—(CH₂)_(n)—CH₃, C₁₋₈-alkyl orC₁₋₈ alkoxy, wherein n and m each independently is an integer in therange of 0 to 3 and wherein at least one of R2, R3, R4, R5 or R6 ishydroxyl, —(CH₂)_(n)—NH—(C═O)—(CH₂)_(n)—CH₃ or —(C═O)—C₁₋₈ haloalkyl;andR7 is as defined herein above in relation to compounds of formula (III).

In this embodiment the 4-oxoquinolizine compounds are preferablycompounds of formula (I) or a pharmaceutically acceptable salt thereof,wherein

R1 is hydrogen or fluorine; andR2, R3, R4, R5 and R6 each independently are hydrogen, hydroxyl,flourine, —NH₂, —(CH₂)—NH₂, C₁₋₃-alkyl, —CH₂—NH—(C═O)—CH₃, —(C═O)—CF₃ orC₁₋₃ alkoxy, wherein at least one of R2,R3, R4, R5 or R6 is hydroxyl, —CH₂—NH—(C═O)—CH₃ or —(C═O)—CF₃; andR7 is C₁₋₃ alkyl or C₁₋₃ alkoxy.

In this embodiment it is even more preferred that the 4-oxoquinolizinecompounds are compounds of formula (I) or a pharmaceutically acceptablesalt thereof, wherein

R1 is hydrogen or fluorine; andR2, R3, R4, R5 and R6 each independently are hydrogen, hydroxyl,—CH₂—NH—(C═O)—CH₃, —(C═O)—CF₃, fluorine, —(CH₂)—NH₂, or methoxy, whereinat least one of R2, R3, R4, R5 or R6 is hydroxyl or —(C═O)—CF₃; andR7 is methyl.

Examples of useful 4-oxoquinolizine compounds according to thisembodiment may for example be selected from the group consisting ofcompounds 56, 61, 63, 64, 66, 70 and 72 of Table 1.

In another embodiment of the invention the preferred 4-oxoquinolizinecompounds are compounds of formula (V) or a pharmaceutically acceptablesalt thereof, wherein

R₁ is hydrogen or fluorine; andR₂, R₃, R₄ and R₅ each individually are selected from the groupconsisting of hydrogen, (CH₂)_(n)-hydroxyl, fluorine, C₁₋₃ alkyl,—(CH₂)_(n)—NH₂, —NH—(CH₂)_(n)—CH₃ and a 5 to 6 membered heterocyclicring, wherein n is an integer in the range of 0 to 2; andR₆ is hydrogen andR₇ is C₁₋₃ alkyl or C₁₋₃ alkoxy; andQ₁, Q₂ and Q₃ each individually are C or N, wherein at least one of Q₁,Q₂ and Q₃ is N and at least one of Q₁, Q₂ and Q₃ is C, and wherein if Q₁is N, then R₃ is not present, and if Q₂ is N, then R₄ is not present andif Q₃ is N, then R₅ is not present.

Preferred compounds of formula (V) such compounds or pharmaceuticallyacceptable salts thereof in which

R₁ is hydrogen or fluorine; andR₂, R₃, R₄ and R₅ each individually are selected from the groupconsisting of hydrogen, hydroxyl, fluorine, methyl, —NH₂, —NH—CH₃ andpiperazinyl; andR₆ is hydrogen andR₇ is methyl or methoxy; andQ₁, Q₂ and Q₃ each individually are C or N, wherein at least one of Q₁,Q₂ and Q₃ is N and at least one of Q₁, Q₂ and Q₃ is C, and wherein if Q₁is N, then R₃ is not present, and if Q₂ is N, then R₄ is not present andif Q₃ is N, then R₅ is not present.

Even more preferred compounds of formula (V) such compounds orpharmaceutically acceptable salts thereof in which

R₁ is hydrogen or fluorine; andR₂ is hydrogen or fluorine; andR₃ is hydrogen or methyl; andR₅ and R₆ are hydrogen andR₇ is methyl; andQ₂ is N, and Q₂ and Q₃ are C, and R₄ is not present.

Other very preferred compounds of formula (V) are such compounds orpharmaceutically acceptable salts thereof in which

R₁ is hydrogen or flourine;R₂ and R₆ are hydrogen; and ′R₃ is hydrogen or methyl; andR₄ is (CH₂)_(n)-hydroxyl, —NH₂ or —NH—CH₃ or a 5 to 6 memberedheterocyclic ring, wherein n is an integer in the range of 0 to 2;preferably R₄ is hydroxyl, —NH₂ or —NH—CH₃ or piperazinyl; andR₇ is methyl or methoxy; andQ₃ is N, and Q₂ and Q₁ are C, and R₅ is not present.

Other very preferred compounds of formula (V) are such compounds orpharmaceutically acceptable salts thereof in which

R₁ is hydrogen or fluorine; andR₂ and R₄ each individually are selected from the group consisting ofhydrogen, (CH₂)_(n)-hydroxyl, fluorine, C₁₋₃ alkyl, —(CH₂)_(n)—NH₂,—NH—(CH₂)_(n)—CH₃ and a 5 to 6 membered heterocyclic ring, wherein n isan integer in the range of 0 to 2; andR₆ is hydrogen; andQ₂ is C, and Q₁ and Q₃ are N, and R₃ and R₅ are not present.

In embodiments of the invention wherein the 4-oxoquinolizine compoundsare compounds of formula (V), then it is preferred that if Q₁ is N andQ₂ and Q₃ are C, then at least one of R₂, R₄ and R₅ is not hydrogen. Inthis embodiment it is also preferred that if Q₁ is N and Q₂ and Q₃ are Cand R₄ is —NH₂, then R₁ is fluorine and/or R₇ is methoxy. Similarly, itis also preferred that if Q₃ is N and Q₂ and Q₁ are C, then at least oneof R₂, R₃ and R₄ is not hydrogen. In this embodiment it is alsopreferred that if Q₃ is N and Q₂ and Q₁ are C and R₄ is —NH₂, then R₁ isfluorine and/or R₇ is methoxy.

Preferred compounds of this embodiment may for example be selected fromthe group consisting of compounds 55, 57, 58, 59, 60, 65, 78, 79, 84, 87and 89 of Table I.

In another embodiment of the invention the 4-oxoquinolizine compoundsare compounds of formula (IIIa) or pharmaceutically acceptable saltsthereof, wherein

R₁ is as defined herein above in relation to compounds of formula (III);andY is a heterobicyclic ring system optionally substituted with one ormore substituents selected from the group consisting of oxo,—(CH₂)_(n)—NH₂, —NH—(CH₂)_(n)—CH₃, —(CH₂)_(n)—OH, C₁₋₈ alkyl, C₁₋₈alkoxy, C₃₋₈-cycloalkyl and halogen, wherein n is an integer in therange of 0 to 3; andR₇ is as defined herein above in relation to compounds of formula (III).

In this embodiment it is preferred that the 4-oxoquinolizine compoundsare compounds of formula (IIIa) or pharmaceutically acceptable saltsthereof, wherein

R₁ is as defined herein above in relation to compounds of formula (III);andY is a 9 membered heterobicyclic ring system, preferably Y is a 9membered heterobicyclic ring system selected from the group consistingof isoindolinyl, indazolyl, benzothiazolyl, benzimidazolyl, indolinyland pyrrolopyridinyl optionally substituted with one or moresubstituents selected from the group consisting of oxo, —(CH₂)_(n)—NH₂,—NH—(CH₂)_(n)—CH₃, —(CH₂)_(n)—OH, C₁₋₈ alkyl, C₁₋₈ alkoxy,C₃₋₈-cycloalkyl and halogen, wherein n is an integer in the range of 0to 3; andR₇ is as defined herein above in relation to compounds of formula (III).

In this embodiment it is preferred that the 4-oxoquinolizine compoundsare compounds of formula (IIIa) or pharmaceutically acceptable saltsthereof, wherein

R₁ is hydrogen, methyl or fluorine; andY is selected from the group consisting of isoindolinyl, indazolyl,benzothiazolyl, benzimidazolyl, indolinyl and pyrrolopyridinyloptionally substituted with one substituent selected from the groupconsisting of oxo, —NH₂, methyl and cyclopropyl; andR₇ methyl or methoxy.

Compounds according to this embodiment may for example be selected fromthe group consisting of compounds 68, 71, 73, 74, 75, 77, 80, 81, 82,83, 85, 86, 88 and 90.

In yet another embodiment of the invention the 4-oxoquinolizinecompounds are compounds of formula (IIIa) or pharmaceutically acceptablesalts thereof, wherein

R₁ is as defined herein above in relation to compounds of formula (III);andY is selected from the group consisting of pyrazolyl andtetrahydropyrinidyl optionally substituted with one or more substituentsselected from the group consisting of oxo, —(CH₂)_(n)—NH₂,—NH—(CH₂)_(n)—CH₃, —(CH₂)_(n)—OH, C₁₋₈ alkyl, C₁₋₈ alkoxy,C₃₋₈-cycloalkyl and halogen, wherein n is an integer in the range of 0to 3; andR₇ is as defined herein above in relation to compounds of formula (III).

In this embodiment it is preferred that the 4-oxoquinolizine compoundsare compounds of formula (IIIa) or pharmaceutically acceptable saltsthereof, wherein

R₁ is hydrogen or flourine; andY is selected from the group consisting of unsubstituted pyrazolyl andtetrahydropyrinidyl; andR₇ is methyl or methoxy.

Compounds of this embodiment may preferably be selected from the groupconsisting of compounds 62 and 67 as mentioned in Table 1.

In specific embodiments, the invention provides the novel antimicrobialcompounds, 1-6 and 17:

In embodiments of the invention relating to 4-oxoquinolizine compoundsused in the absence of polymyxins it is preferred that said4-oxoquinolizine compounds are not any of the compounds described inExamples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106,107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120,121, 122 and 123 of US2004/0229903.

In embodiments of the invention relating to 4-oxoquinolizine compoundsused in the absence of polymyxins it is preferred that said4-oxoquinolizine compounds are not any of compounds 1, 2, 3, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, 50, 51, 52, 53 or 54 of Table 1.

In embodiments of the invention relating to 4-oxoquinolizine compoundsused in the absence of polymyxins it is preferred that said4-oxoquinolizine compounds are not any of compounds named Examples 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 or 52 of PCT applicationPCT/US2011/052003.

Salts of 4-Oxoquinolizines

Pharmaceutically acceptable salts of the 4-oxoquinolizine compounds ofthe invention may include acid or base addition salts. Acid and baseaddition salts refers to the relatively non-toxic, inorganic and organicaddition salts of compounds of the present invention. These salts can beprepared in situ during the final isolation and purification of thecompounds, or by subsequently reacting the purified compound in its freeacid or base form with a suitable organic or inorganic compound andisolating the salt thus formed. In so far as the compounds of formula(I) of this invention are basic compounds, they are all capable offorming a wide variety of different salts with various inorganic andorganic acids. Although such salts must be pharmaceutically acceptablefor administration to animals, it is often desirable in practice toinitially isolate the base compound from the reaction mixture as apharmaceutically unacceptable salt and then simply convert to the freebase compound by treatment with an alkaline reagent and thereafterconvert the free base to a pharmaceutically acceptable acid additionsalt.

The pharmaceutically acceptable acid addition salts of the basiccompounds are prepared by contacting the free base form with asufficient amount of the desired acid to produce the salt in theconventional manner. The free base form may be regenerated by contactingthe salt form with a base and isolating the free base in theconventional manner. The free base forms differ from their respectivesalt forms somewhat in certain physical properties such as solubility inpolar solvents, but otherwise the salts are equivalent to theirrespective free base for purposes of the present invention.

Pharmaceutically acceptable base addition salts are formed with metalsor amines. The base addition salts of acidic compounds are prepared bycontacting the free acid form with a sufficient amount of the desiredbase to produce the salt in the conventional manner. The free acid formmay be regenerated by contacting the salt form with an acid andisolating the free acid in a conventional manner. The free acid formsdiffer from their respective salt forms somewhat in certain physicalproperties such as solubility in polar solvents, but otherwise the saltsare equivalent to their respective free acid for purposes of the presentinvention.

Salts may be prepared from inorganic acids or organic acids. One exampleof a salt of the 4-oxoquinolizines according to the invention may behydrochloric salts.

Another example of a salt of the 4-oxoquinolizines according to theinvention is potassium salts.

Methods of Preparing 4-Oxoquinolizines and Salts Thereof

The 4-oxoquinolizine compounds of the invention described herein in thesections “4-oxoquinolizines” and “Particular 4-oxoquinolizines” may beprepared using the methods outlined herein below in Example 8. Inparticular, useful scaffolds of the 4-oxoquinolizine compounds may beprepared as described in Example 8 and these may be further modifiedusing methods known to the skilled person. Thus, suitable scaffolds maybe prepared in 6 to 7 steps as described in Example 8. The suitablescaffolds may then be used to obtain the final 4-oxoquinolizinecompounds in 2 to 5 steps.

The 4-oxoquinolizine compounds of the invention may also be prepared asdescribed for specific 4-oxoquinolizine compounds in Example 8. Thesemethods may optionally be modified with relevant modifications known tothe skilled person.

Example 8 also described useful examples of preparing salts of4-oxoquinolizine compounds. These methods may also be modified by theskilled person to prepare salts of other 4-oxoquinolizines.

The 2 pyridones to be used in pharmaceutical compositions comprisingboth a 4-oxoquinolizine compound and a Polymyxin may also be prepared asdescribed in US patent application US2004/0229903.

Properties of Preferred 4-Oxoquinolizines

The preferred 4-oxoquinolizines according to the invention arepreferably active antimicrobial compounds.

In particular the 2-pyridone compounds, such as the 4-oxoquinolizinecompounds of the invention show potent activity against a pathogen thatis resistant against one or more of quinolones, carbapenems, aminosidesand glycopeptides antibiotics, and specifically, against a pathogen thatis a CDC category A or category B pathogen. The 4-oxoquinolizinecompounds may in particular have a MIC≦10 μg/ml, such as ≦5 μg/ml, forexample ≦1 μg/ml, such as ≦0.5 μg/ml against B. thailandensis whendetermined as described herein below in Example 1. B. thailandensis is asurrogate strain, which may be used experimentally instead of B.pseudomallei. Thus, the 4-oxoquinolizine compounds may have a MIC≦10μg/ml, such as ≦5 μg/ml, for example ≦1 μg/ml, such as ≦0.5 μg/mlagainst one or more of, preferably against all of B. anthracis, F.tularensis, B. acortus and B. pseudomallei when determined as describedherein below in Example 1.

The 4-oxoquinolizine compounds of the invention may also have a MIC≦10μg/ml, such as 55 μg/ml against one or more bacteria selected from thegroup consisting of S. aureus, S. epidermis, E. faecalis and E. faeciumwhen determined as described herein below in Example 2.

The 4-oxoquinolizine compounds of the invention may also have a MIC≦50μg/ml, such as 520 μg/ml against A. baumanni, wherein said A. baumanniis a drug resistant strain of A. baumanni, for example a multidrugresistant strain of A. baumanni when determined as described hereinbelow in Example 3.

The 4-oxoquinolizine compounds of the invention preferably exhibitpotent activities in presence of subinhibitory concentrations ofpolymyxin B against A. baumannii including quinolone and multiresistantstrains of A. baumannii. The antimicrobial activity of the4-oxoquinolizine compounds when combined with subinhibitoryconcentrations of polymyxin B against Acinetobacter baumannii includingmultiresistant clinical isolates may be determined as described hereinbelow in Example 4. Thus the 4-oxoquinolizine compounds of the inventionmay have a MIC≦20 μg/ml, such as ≦15 μg/ml, for example ≦10 μg/ml, suchas ≦5 μ/ml against A. baumannii in the presence of 0.06 μg/ml PolymyxinB when determined as described in Example 4 herein below.

The particular 4-oxoquinolizine compounds of the invention may also havea MIC≦10 μg/ml, such as ≦5 μg/ml, for example 1lμg/ml, such as ≦0.5μg/ml against at least one Gram negative bacterium and against at leastone gram positive bacterium when determined as described herein below inExample 5.

The 4-oxoquinolizine compounds of the invention in general have at leastpartly synergistic antibacterial activity with Polymyxin, such as withPolymyxin B. This may for example be determined as described hereinbelow in example 6. Thus, the 4-oxoquinolizine compounds of theinvention may have a FICI<0.75, such as ≦0.5 in relation to Polymyxin Bagainst A. baumannii and/or K. pneumoniae, when determined as describedherein below in Example 6.

The 4-oxoquinolizine compounds of the invention preferably have a lowcytotoxicity against human cells. Thus the 4-oxoquinolizine compounds ofthe invention may have an IC50 of at least 20 μM, such as at least 40 μMin respect of HEPG2 V1 cells when determined as described herein belowin Example 7.

Polymyxin

It is an aspect of the present invention to provide pharmaceuticalcompositions comprising a 4-oxoquinolizine compound in combination witha Polymyxin. The Polymyxin to be used with the present invention may beany useful Polymyxin.

Polymyxins are antibiotics isolated from cultures of Bacillus polymyxa.In general Polymyxins are active as antibiotics against mostGram-negative bacteria. Polymyxins according to the present inventionare preferably decapeptides containing five or six residues ofL-2,4-diaminobutyric acid. Preferably, the sequence of seven residues atthe C-terminal end of the decapeptide is formed into a peptide ringthrough an isopeptide link to one of the diaminobutyric acid residues,while the N-terminal residue preferably is acylated with a fatty acid.The fatty acid is preferably —(CH₂)_(m)—COOH, wherein m is an integer inthe range of 6 to 15, preferably in the range of 7 to 10, morepreferably in the range of 7 to 8, wherein said —(CH₂)_(m)—COOHoptionally may be substituted with one or more C₁₋₈ alkyl, preferablywith one C₁₋₈ alkyl, more preferably with one C₁₋₅ alkyl, even morepreferably with one C₁₋₂ alkyl, yet more preferably with one methyl.

Preferred Polymixins to be used with the present invention are PolymyxinB or Polymyxin E or pharmaceutically acceptable salts thereof.

Polymyxin B is a lipopeptide antibiotic originally isolated fromBacillus polymyxa. Polymyxin B according to the present inventionpreferably consists of a peptide ring of 7 amino acids and a tripeptideside chain with a fatty acid tail. Polymyxin B according to theinvention preferably contains five primary amine groups and is thus apolycation at pH 7.

Polymyxin B to be used with the present invention may be selected fromthe group consisting of Polymyxin B1 Polymyxin B2, Polymyxin B3 andPolymyxin B4. Polymyxin B to be used with the present invention may alsobe a mixture of 2 or more of the aforementioned, preferably Polymyxin Bis a mixture containing at least Polymyxin B1 and Polymyxin B2, morepreferably Polymyxin B to be used with the present invention is amixture containing Polymyxin B1 Polymyxin B2, Polymyxin B3 and Polymyxin84, Polymyxin B according to the present invention is preferably acompound of the formula (V)

or a pharmaceutically acceptable salt thereof, wherein Dab is2,4-diaminobutyric acid; and α and γ indicate which amino group isinvolved in the peptide linkage. The 3-letter code for amino acids isused.

In relation to formula (V) the fatty acid is preferably selected fromthe group consisting of heptanoic acid and octanoic acid, optionallysubstituted with methyl,

Preferably Polymyxin B1 is a compound of formula (V) wherein the fattyacid is 6-methyloctanoic acid. Preferably Polymyxin B2 is a compound offormula (V) wherein the fatty acid is 6-methylheptanoic acid. PreferablyPolymyxin B3 is a compound of formula (V) wherein the fatty acid isoctanoic acid. Preferably Polymyxin B4 is a compound of formula (V)wherein the fatty acid is heptanoic acid.

Thus, Polymyxin B1 and B2 may preferably be compounds of the formula(VI)

wherein R is —H in Polymyxin B1, and R is —CH₃ in polymyxin B2.

The pharmaceutically acceptable salt of Polymyxin B may preferably beany of the salts prepared from an inorganic acid mentioned herein abovein the section “Salts of 4-oxoquinolizines”, a more preferably said saltis the sulfate salt.

The Polymyxin to be used with the present invention may in embodiment bePolymyxin E, which may also be referred to as colistin. Polymyxin E maypreferably be a compound of the formula (VI):

or a pharmaceutically acceptable salt thereof. Polymyxin E may in oneembodiment be available as a prodrug, more preferably as colistinmethanesulphonate.

The pharmaceutically acceptable salt of Polymyxin E may for example bethe sodium salt, such as the sodium salt of colistin methanesulphonate.

Dosage of Polymyxin

It is one aspect of the present invention to provide pharmaceuticalcompositions comprising a 4-oxoquinolizine compound in combination withPolymyxin, preferably Polymyxin B. In one embodiment of the inventionsaid Polymyxin, e.g. Polymyxin B is present in a subinhibitoryconcentration.

Said subinhibitory concentration is in general dependent on theindividual to receive treatment with the pharmaceutical composition,Preferably said subinhibitory concentration of Polymyxin isadministration of a less than 2 mg Polymyxin per kg of said individualper day, Thus the subinhibitory concentration may be administration ofless than 1.5 mg, such as less than 1 mg, for example less than 0.5 mg,such as less than 0.3 mg, for example less than 0.1 mg per kg of saidindividual per day. Preferably said subinhibitory concentration ofPolymyxin B is administration of a less than 2 mg Polymyxin B per kg ofsaid individual per day, Thus the subinhibitory concentration may beadministration of less than 1.5 mg, such as less than 1 mg, for exampleless than 0.5 mg, such as less than 0.3 mg, for example less than 0.1 mgPolymyxin B per kg of said individual per day.

Thus, in embodiments of the invention where the pharmaceuticalcomposition is prepared in daily unit dosages for administration toadult human being, then each dosage unit preferably comprises less than150 mg Polymyxin, such as less than 130 mg Polymyxin, for example lessthan 110 mg Polymyxin, such as less than 90 mg Polymyxin, for exampleless than 70 mg Polymyxin, such as less than 50 mg Polymyxin, whereinthe Polymyxin may be any of the Polymyxins described herein above in thesection “Polymyxin”, but preferably is Polymyxin B.

In one embodiment of the invention the subinhibitory concentration isless than 0.5 μg/ml, such as less than 0.4 μg/ml, for example less than0.3 μg/ml, such as less than 0.2 μg/ml.

Polymyxin may however also be administered at regular concentrations,for example in the range of 2 to 5 mg Polymyxin, such as Polymyxin B perkg of said individual may be administered.

Compositions Comprising Polymyxin and a 4-Oxoquinolizine Compound

It is one aspect of the invention to provide pharmaceutical compositionscomprising

-   -   a) A Polymyxin, which may be any of the Polymyxins described        herein above in the section “Polymyxin”; and    -   b) A 4-oxoquinolizine compound, which may be any of the        4-oxoquinolizine compounds described herein above in the section        “4-oxoquinolizine compound” or the section “Particular        4-oxoquinolizine compound”.

Interestingly, the present invention discloses that the antibacterialeffect of Polymyxin and 4-oxoquinolizines is synergistic. Thesynergistic effect may be determined according to any useful method,such as using a checkerboard technique, e.g. by using the methoddescribed in Example 6 herein below.

Thus, in one embodiment of the invention the compositions comprises asubinhibitory concentration of a polymyxin, for example Polymyxin B anda subinhibitory concentration of a 4-oxoquinolizine, wherein saidcomposition is capable of inhibiting growth of at least one bacterium,more preferably the composition is capable of inhibiting growth of atleast 2 different bacteria, for example of at least 5 differentbacteria, such as of at least 10 different bacteria.

In one embodiment of the invention the pharmaceutical compositionscomprises

-   -   a) Polymyxin B; and    -   b) a 4-oxoquinolizine compound, which may be any of the        4-oxoquinolizine compounds described herein above in the section        “4-oxoquinolizine compound” or the section “Particular        4-oxoquinolizine compound”, wherein the 4-oxoquinolizine        compound has an antibacterial effect against at least one        bacterium, such as against at least 2 different bacteria, for        example against at least 5 different bacteria, such as against        at least 10 different bacteria which is synergistic with the        antibacterial effect of polymyxin B, wherein the synergistic        effect preferably is determined as described herein below in        Example 6.

In one aspect the invention relates to a method of treating a bacterialinfection in an individual in need thereof, wherein the method comprisesthe steps of:

-   -   i) administering Polymyxin B to a individual in need thereof;        and    -   ii) testing whether the bacterial infection is reduced or cured        by said Polymyxin B; and    -   iii) if the bacterial infection is not reduced or cured by said        Polymyxin B, then administering to said individual a        therapeutically effective amount of a 4-oxoquinolizine compound,        which may be any of the 4-oxoquinolizine compounds described        herein above in the section “4-oxoquinolizine compound” or the        section “Particular 4-oxoquinolizine compound”;    -   thereby treating said bacterial infection in said individual.

The Polyxin may be administered in any useful dosage, such as any of thedosages described herein above in the section “Dosage of Polymyxin”. Forexample the polymyxin may be administered at in the range of 2 to 5 mgPolymyxin, such as Polymyxin B per kg of said individual, or thepolymyxin may even be administered at subinhibitory concentrations.

The 4-oxoquinolizine compound may preferably be administered asdescribed herein below in the section “Pharmaceutical formulations”.

Bacterial Infection

It is an aspect of the present invention to provide pharmaceuticalcompositions comprising a 4-oxoquinolizine compound in combination witha Polymyxin. Said pharmaceutical compositions are in particular usefulfor treatment of a bacterial infection in an individual in need thereof,and preferably for treatment of any of the bacterial infectionsdescribed herein in this section.

It is furthermore an aspect of the present invention to provideparticularly useful 4-oxoquinolizine compounds as described herein abovein the section “Particular 4-oxoquinolizines”. Said 4-oxoquinolizinecompounds are in particular useful for treatment of a bacterialinfection in an individual in need thereof, and preferably for treatmentof any of the bacterial infections described herein in this section.

In general the pharmaceutical compositions comprising a 4-oxoquinolizinecompound in combination with a Polymyxin or the particular4-oxoquinolizine compounds are invention are useful for treatment of abroad spectrum of different bacterial infections. Thus, they aretypically useful for treatment infections by at least two differentkinds of bacteria, such as at least 5 different kinds of bacteria, forexample at least 10 different kinds of bacteria. In particular, saidbacteria may be drug resistant bacteria, such as multidrug resistantbacteria.

The 4-oxoquinolizine compounds and the pharmaceutical compositionscomprising 4-oxoquinolizine and Polymyxin are antibacterial agentsusable for the treatment of local infectious diseases or generalinfectious diseases of human beings or animals caused by Gram-positivebacteria, Gram-negative bacteria, anaerobic bacteria, acid-fast bacteriaor other bacteria.

This invention also provides methods of treating an infectious disorderin an individual in need thereof, wherein the individual for example maybe a human or a mammal, by administering a safe and effective amount ofa 4-oxoquinolizine compound to said subject optionally in combinationwith a Polymyxin. As used herein, an “infectious disorder” is anydisorder characterized by the presence of a microbial infection.Exemplary methods of this invention are for the treatment of bacterialinfections. Such infectious disorders include for example centralnervous system infections, external ear infections, infections of themiddle ear (such as acute otitis media), infections of the cranialsinuses, eye infections, infections of the oral cavity (such asinfections of the teeth, gums and mucosa), upper respiratory tractinfections, lower respiratory tract infections, including pneumonia,genitourinary infections, gastrointestinal infections, gynecologicalinfections, septicemia, sepsis, peritonitis, bone and joint infections,skin and skin structure infections, bacterial endocarditis, burns,antibacterial prophylaxis of surgery, or antibacterial prophylaxis inpost-operative patients or in immunosuppressed patients (such aspatients receiving cancer chemotherapy, or organ transplant patients).

Thus the bacterial infection may be infection by one or more bacteria offor example the central nervous system, external ear, the middle ear(such as acute otitis media), the cranial sinuses, the eyes, the oralcavity (such as of the teeth, gums and mucosa), upper respiratory tract,lower respiratory tract, including lung, genitourinary tract,gastrointestinal tract, peritoneum, bone and joints, skin or burns. Thebacterial infection may also be related to sepsis, surgery, or bacterialinfecitons in post-operative patients or in immunosuppressed patients.

The bacterial infection may be infection by any bacteria; preferably thebacterium is pathogenic bacterium. The bacterial infection may beinfection by a gram-negative or a gram-positive bacterium or it may beinfection by a mixture of bacteria, which may be gram-positive orgram-negative. The present invention discloses that 4-oxoquinolizinecompounds are effective in treatment of infections by both Gram-positiveand Gram-negative bacteria.

In embodiments of the invention relating to pharmaceutical compositionscomprising both 4-oxoquinolizine compounds and a Polymyxin, then it ispreferred that the bacteria is a Gram-negative bacterium.

The bacterial infection to be treated with the pharmaceuticalcomposition comprising a 4-oxoquinolizine compound and a Polymyxin orwith the particular 4-oxoquinolizine compound according to the presentinvention is preferably infection by one or more bacteria of a genusselected from the group consisting of:

Acinetobacter, Bacillus, Bortadella, Borrelia, Brucella, Camphylobacter,Chlamydia, Clostridium, Corynebacterium, Enterococcus, Escherichia,Fransisella, Haemophilus, Helicobacter, Legionella, Leptospira,Listeria, Mycobacterium, Mycoplasma, Neisseria, Propionibacterium,Pseudomonas, Rickettsia, Salmonella, Shigella, Staphylococcus,Streptococcus, Treponema, Vibrio and Yersinia.

The bacterial infection to be treated with the pharmaceuticalcomposition comprising a 4-oxoquinolizine compound and a Polymyxin orwith the particular 4-oxoquinolizine compound according to the presentinvention is preferably infection by one or more bacteria of genusselected from the group consisting of:

Acinetobacter, Bacillus, Brucella, Burkholderia, Citrobacter,Corynebacterium, Enterobacter, Enterococcus, Escherichia, Francisella,Haemophilus, Klepsiella, Listeria, Moraxella, Morganella, Neisseria,Proteus, Providencia, Pseudomonas, Serratia, Shigella, Staphylococcus,Stenotrophomonas, Streptococcus and Yersinia.

In one embodiment of the invention the bacterial infection to be treatedwith the pharmaceutical composition comprising a 4-oxoquinolizinecompound and a Polymyxin or with the particular 4-oxoquinolizinecompound according to the present invention is infection by one or morebacteria of a genus selected from the group consisting of Acinetobacter,Bortadella, Borrelia, Brucella, Camphylobacter, Chlamydia, Clostridium,Corynebacterium, Fransisella, Helicobacter, Legionella, Leptospira,Listeria, Mycobacterium, Mycoplasma, Neisseria, Rickettsia, Salmonella,Shigella, Treponema, Vibrio and Yersinia.

The bacterial infection to be treated with the pharmaceuticalcomposition comprising a 4-oxoquinolizine compound and a Polymyxin orwith the particular 4-oxoquinolizine compound according to the presentinvention is preferably infection by one or more bacteria of a genusselected from the group consisting of:

Acinetobacter, Brucella, Burkholderia, Citrobacter, Corynebacterium,Enterobacter, Francisella, Listeria, Moraxella, Morganella, Neisseria,Proteus, Providencia, Serratia, Shigella, Stenotrophomonas, andYersinia.

The bacterial infection to be treated with the pharmaceuticalcomposition comprising a 4-oxoquinolizine compound and a Polymyxin orwith the particular 4-oxoquinolizine compound according to the presentinvention is preferably infection by one or more bacteria selected fromthe group consisting of:

Acinetobacter spp, Acinetobacter baumannii, Bacillus anthracis, Brucellaabortus, Burkholderia cepacia, Burkholderia mallei, Burkholderiapseudomallei, Burkholderia thailandensis, Citrobacter freundii,Corynebacterium jeikeium, Enterobacter sp, Enterobacter cloacae,Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum,Escherichia coli, Francisella tularensis, Haemophilus influenza,Klepsiella spp., Klebsiella aerogenes, Klebsiella pneumoniae, Listeriamonocytogenes, Moraxella catarrhalis, Morganella morganii, Neisseriameningitides, Proteus mirabilis, Providencia stuartii, Pseudomonas spp.,Pseudomonas aeruginosa, Serratia marcescens, Shigella sp, Staphylococcusaureus, Staphylococcus epidermis, Staphylococcus haemolyticus,Staphylococcus saprophyticus, Stenotrophomonas maltophilia,Streptococcus agalactiae, Streptococcus bovis, Streptococcusconstellatus, Streptococcus mitis, Streptococcus pneumoniae,Streptococcus pyogenes, Streptococcus oralis, Streptococcus sanguis,Group C Streptococcus, Yersinia pestis, and drug-resistant strainsthereof.

In an embodiment of any of the preceding embodiments, the bacteria canbe selected from the group consisting of, (a) Acinetobacter baumannii,Bacillus anthracis, Brucella abortus, Burkholderia cepacia, Burkholderiamallei, Burkholderia pseudomallei, Burkholderia thailandensis,Citrobacter freundii, Corynebacterium jeikeium, Enterobacter sp,Enterobacter cloacae, Enterococcus faecium, Enterococcus gallinarum,Francisella tularensis, Klebsiella aerogenes, Listeria monocytogenes,Moraxella catarrhalis, Morganella morganii, Neisseria meningitides,Proteus mirabilis, Providencia stuartii, Serratia marcescens, Shigellasp, Staphylococcus haemolyticus, Staphylococcus saprophyticus,Stenotrophomonas maltophilia, Streptococcus agalactiae, Streptococcusbovis, Streptococcus constellatus, Streptococcus mitis, Streptococcuspyogenes, Streptococcus oralis, Streptococcus sanguis, Group CStreptococcus, and Yersinia pestis, (b) drug-resistant strains of anypathogen of part (a); and (c) and drug-resistant strains ofStaphylococcus aureus, Staphylococcus epidermis, Streptococcuspneumoniae, Enterococcus faecalis, Escherichia coli, Klebsiellapneumoniae, Pseudomonas aeruginosa, and Haemophilus influenza.

By the term “drug-resistant strain” is meant a bacterial strain which isresistant to at least one antibiotic drug selected from the groupconsisting of quinolones, carbapenems, aminosides and glycopeptidesantibiotics. Multidrug-resistant strains are bacterial strains resistantto at least two two antibiotic drugs of different classes, wherein saidclasses are selected from the group consisting of quinolones,carbapenems, aminosides and glycopeptides antibiotics.

The bacterial infection to be treated with the 4-oxoquinolizinecompounds described in the section “Particular 4-oxoquinolizines” arepreferably bacteria selected from the group consisting of:

Acinetobacter spp, Acinetobacter baumannii, Bacillus anthracis, Brucellaabortus, Burkholderia cepacia, Burkholderia mallei, Burkholderiapseudomallei, Burkholderia thailandensis, Citrobacter freundii,Corynebacterium jeikeium, Enterobacter sp, Enterobacter cloacae,Enterococcus gallinarum, Francisella tularensis, Klebsiella aerogenes,Listeria monocytogenes, Moraxella catarrhalis, Morganella morganii,Neisseria meningitides, Proteus mirabilis, Providencia stuartii,Serratia marcescens, Shigella sp, Staphylococcus haemolyticus,Staphylococcus saprophyticus, Stenotrophomonas maltophilia,Streptococcus agalactiae, Streptococcus bovis, Streptococcusconstellatus, Streptococcus mitis, Streptococcus pyogenes, Streptococcusoralis, Streptococcus sanguis, Group C Streptococcus, Yersinia pestis,and drug-resistant strains thereof.

The bacterial infection to be treated with the pharmaceuticalcomposition comprising a 4-oxoquinolizine compound and a Polymyxin orwith the particular 4-oxoquinolizine compound according to the presentinvention may be selected from the group consisting of Burkholderiapseudomallei, Bacillus anthracis, Yersinia pestis, Francisellatularensis, and Brucella abortu, Klebsiella, Pseudomonas, Acinetobacterand Staphylococcus aureus, wherein said Staphylococcus aureus may beMethicillin-resistant Staphylococcus aureus (MRSA).

In one embodiment the bacterial infection to be treated with thepharmaceutical composition comprising a 4-oxoquinolizine compound and aPolymyxin or with the particular 4-oxoquinolizine compound according tothe present invention the infection is infection by a Gram-positivebacterium. This may in particular be the case in embodiments of theinvention relating to the particular 4-oxoquinolizine compoundsdescribed herein above in the section “Particular 4-oxoquinolizines”.Said Gram-positive bacterium may for example be selected from the groupconsisting of B. anthracis, S. epidermidis, Staphylococcus aureus,Streptococcus aureus, Streptococcus pneumonia, Enterococcus faecalis andEnterococcus faecium.

The bacterial infection to be treated with pharmaceutical compositionscomprising 4-oxoquinolizine compounds and Polymyxins may preferably bebacteria of a genus selected from the group consisting of:

Acinetobacter, Bortadella, Borrelia, Brucella, Camphylobacter,Escherichia, Fransisella, Haemophilus, Helicobacter, Legionella,Leptospira, Neisseria, Pseudomonas, Rickettsia, Salmonella, Shigella,Treponema, Vibrio and Yersinia.

The bacterial infection to be treated with pharmaceutical compositionscomprising 4-oxoquinolizine compounds and Polymyxins may preferably beselected from bacteria of a genus selected from the group consisting of

Acinetobacter, Brucella, Burkholderia, Citrobacter, Enterobacter,Escherichia, Francisella, Haemophilus, Klepsiella, Moraxella,Morganella, Neisseria, Proteus, Providencia, Pseudomonas, Serratia,Shigella, Stenotrophomonas and Yersinia.

In one embodiment the bacterial infection to be treated with thepharmaceutical composition comprising a 4-oxoquinolizine compound and aPolymyxin or with the particular 4-oxoquinolizine compound according tothe present invention the infection is infection by a Gram-negativebacterium. Said Gram-negative bacterium may for example be selected fromthe group consisting of F. tularensis, B. abortus, B. pseudomallei,Pseudomonas aeruginosa, Burkholderia thailandensi, Acinetobacterbaumannii, Acinetobacter Escherichia coli and Klebsiella.

In one embodiment the bacterial infection to be treated with thepharmaceutical composition comprising a 4-oxoquinolizine compound and aPolymyxin or with the particular 4-oxoquinolizine compound according tothe present invention the infection is infection by an anaerobicbacterium. Said anaerobic bacterium may for example be Bacillusfragilis.

In particular the bacteria may be a bacterial strain that is resistantagainst one or more of quinolones, carbapenems, aminosides andglycopeptides antibiotics, and specifically, the bacterial infection maybe infection by a pathogen that is a CDC category A or category Bpathogen. CDC is the US Centers for Disease Control and Prevention,which have categorised pathogens according to risk to national security(Example 1 below describe examples of such bacteria).

Thus, the bacterial infection to be treated with the 4-oxoquinolizinecompounds of the invention or with pharmaceutical compostions comprising4-oxoquinolizine compounds and Polymyxin according to the presentinvention may in one embodiment be a Category A bacteria according toCDC's classification. Category A bacteria are defined as bacteria that

-   -   can be easily disseminated or transmitted from person to person;    -   result in high mortality rates and have the potential for major        public health impact;    -   might cause public panic and social disruption; and    -   require special action for public health preparedness.

Category A bacteria may for example be selected from the groupconsisting of Bacillus anthracis, Clostridium botulinum, Yersinia pestisand Francisella tularensis.

Thus, the bacterial infection to be treated with the 4-oxoquinolizinecompounds of the invention or with pharmaceutical compostions comprising4-oxoquinolizine compounds and Polymyxin according to the presentinvention may in one embodiment be a Category B bacteria according toCDC's classification. Category B bacteria are defined as bacteria that

-   -   are moderately easy to disseminate;    -   result in moderate morbidity rates and low mortality rates; and    -   require specific enhancements of CDC's diagnostic capacity and        enhanced disease surveillance.

Category B bacteria may for example be selected from the groupconsisting of Brucella species, Clostridium perfringens, Salmonellaspecies, Escherichia coli O157:H7, Shigella, B. abortus, Burkholderiamallei, Burkholderia pseudomallei, Chlamydia psittaci, Coxiellaburnetii, Staphylococcus spp. Rickettsia prowazekii, Vibrio cholerae andCryptosporidium parvum.

The term “treatment” is used herein to mean that, at a minimum,administration of a compound of the present invention mitigates adisease associated an infectious disorder, e.g. a bacterial infection ina host, such as in a mammalian subject, such as in humans. Thus, theterm “treatment” includes inhibiting the infectious disorder; and/oralleviating or reversing the infectious disorder. Insofar as the methodsof the present invention are directed to preventing infectiousdisorders, it is understood that the term “prevent” does not requirethat the disease state be completely thwarted. (See Webster's NinthCollegiate Dictionary.) Rather, as used herein, the term preventingrefers to the ability of the skilled artisan to identify a populationthat is susceptible to infectious disorders, such that administration ofthe compounds of the present invention may occur prior to onset ofinfection. The term does not imply that the disease state be completelyavoided.

Pharmaceutical Formulations

The present invention relates in one aspect to pharmaceuticalcompositions comprising 4-oxoquinolizine compounds and Polymyxin. Thepresent invention also relates to pharmaceutical compositions comprisingpreferred 4-oxoquinolizine compounds with or without the presence ofPolymyxins.

According to the present invention, the pharmaceutical compositions arepreferably for treatment of an individual infected by the pathogen, suchas an individual suffering from a bacterial infection. The compositionsmay however also be for administration to an individual at risk ofacquiring such an infection. Generally, the individual is a vertebrate,preferably a mammal, and more preferably a human being. The treatmentmay be ameliorating or curative. By curative, it is intended to meansurvival from the infection which otherwise in the absence of thetreatment causes the subject suffering from the infection to showincreasing pathology or even morbidity. Thus, the pharmaceuticalcompositions described herein may be prepared for prophylacticadministration to an individual at risk of infection by the pathogen,preferably by the bacteria.

In one embodiment of the invention, the pharmaceutical composition isfor reducing the risk of contagion caused by the infection or in anindividual at risk of acquiring a bacterial infection. In relation toepidemic or even pandemic infections causing a high mortality rate, evenslight reductions in risk of contagion may be of major importance.

In another embodiment, the pharmaceutical composition reduces the riskof contagion in a individual that has acquired a bacterial infection byat least 5%, preferably at least 10%, preferably at least 15%, morepreferably at least 20%, or at least 30% or at least 40%, or at least50%, or at least 60%, or at least 70% or at least 80%, or at least 90%,or more. The pharmaceutical compositions of the invention may alsoreduce the risk of contagion caused by a bacterial infection in anindividual at risk of acquiring that infection by at least 5%, or atleast 10%, or at least 15%, or at least 20%, or at least 30% or at least40%, or at least 50%, or at least 60%, or at least 70% or at least 80%,or at least 90%, or more.

Administration of the anti-bacterial pharmaceutical compositionsaccording to the invention may be only once or administration may berepeated for a number of times. For example, the pharmaceuticalcompositions comprising 4-oxoquinolizine compounds or the pharmaceuticalcompositions comprising both Polymyxins and 4-oxoquinolizine compoundsmay be given repeatedly with regular intervals, for example in the rangeof 1 to 5 times daily for in the range of 1 to 100 days, such as in therange of 1 to 50 days, for example in the range of 1 to 25 days, such asin the range of 10 to 16 days. The total daily dose of the compounds ofthis invention administered to a host in single or in divided doses canbe in amounts, Single dose compositions may contain such amounts orsubmultiples thereof as make up the daily dose. Preferred dosages ofPolymyxin to be contained in these compositions are described herein inthe section “Dosage of Polymyxin” and preferred dosages of4-oxoquinolizine compounds are described herein below.

The pharmaceutical compositions may be prepared for any suitableadministration route, for example, topical, parenteral, oral, buccal,systemic, nasal, injection, transdermal, rectal, vaginal, etc., or aform suitable for administration by inhalation or insufflation. Forexample, the pharmaceutical compositions of the invention are preparedfor oral administration or for intraperitoneal administration, such asfor oral administration. Similarly, the pharmaceutical compositions ofthe invention may or may be used at the site of a wound on or in thebody, for example as a result of surgery or injury. Equally, thepharmaceutical compositions of the invention may or may be used for aninternal infection at the site of a prosthesis.

Formulations suitable for oral administration can consist of (a) liquidsolutions, such as an effective amount of the active compound(s)suspended in diluents, such as water, saline or PEG 400; (b) capsules,sachets or tablets, each containing a predetermined amount of the activeingredient, as liquids, solids, granules or gelatin; (c) suspensions inan appropriate liquid; and (d) suitable emulsions. Tablet forms caninclude one or more of lactose, sucrose, mannitol, sorbitol, calciumphosphates, corn starch, potato starch, microcrystalline cellulose,gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearicacid, and other excipients, colorants, fillers, binders, diluents,buffering agents, moistening agents, preservatives, flavoring agents,dyes, disintegrating agents, and pharmaceutically compatible carriers.

Lozenge forms can comprise the active ingredient(s) in a flavor, e.g.,sucrose, as well as pastilles comprising the active ingredient in aninert base, such as gelatin and glycerin or sucrose and acaciaemulsions, gels, and the like containing, in addition to the activeingredient(s), carriers known in the art.

The 4-oxoquinolizine compounds or the compositions comprising bothPolymyxins and 4-oxoquinolizine compounds as described above, alone orin combination with other suitable components, can be made into aerosolformulations (i.e., they can be “nebulized”) to be administered viainhalation. Aerosol formulations can be placed into pressurizedacceptable propellants, such as dichlorodifluoromethane, propane,nitrogen, and the like.

Suitable formulations for rectal administration include, for example,suppositories, which consist of the packaged compound with a suppositorybase. Suitable suppository bases include natural or synthetictriglycerides or paraffin hydrocarbons. In addition, it is also possibleto use gelatin rectal capsules which consist of a combination of thecompound of choice with a base, including, for example, liquidtriglycerides, polyethylene glycols, and paraffin hydrocarbons.

Formulations suitable for parenteral administration, such as, forexample, by intraarticular (in the joints), intravenous, intramuscular,intradermal, intraperitoneal, and subcutaneous routes, include aqueousand non-aqueous, isotonic sterile injection solutions, which can containantioxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.In the practice of this invention, compositions can be administered, forexample, by intravenous infusion, orally, topically, intraperitoneally,intravesically or intrathecally.

In addition, dosages for injection of the pharmaceutical compositions ofthe invention may be prepared in dried or lyophilized form. Such formscan be reconstituted with water or saline solution, depending on thepreparation of the dosage form. Such forms may be packaged as individualdosages or multiple dosages for easier handling. Where lyophilized ordried dosages are used, the reconstituted dosage form may be isotonic,and at a physiologically compatible pH.

Various oral dosage forms of the pharmaceutical compositions of theinvention can be used, including such solid forms as tablets, capsules,granules and bulk powders. These oral forms comprise a safe andeffective amount, usually at least about 5%, or from about 25% to about50%, of the 4-oxoquinolizine compounds and optionally they may alsocomprise Polymyxins. Tablets can be compressed, tablet triturates,enteric-coated, sugar-coated, film-coated, or multiple-compressed,containing suitable binders, lubricants, diluents, disintegratingagents, coloring agents, flavoring agents, flow-inducing agents, andmelting agents. Liquid oral dosage forms include aqueous solutions,emulsions, suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules, and effervescent preparations reconstitutedfrom effervescent granules, containing suitable solvents, preservatives,emulsifying agents, suspending agents, diluents, sweeteners, meltingagents, coloring agents and flavoring agents, such are well known to theskilled artisan. Exemplary excipients for oral administration includegelatin, propylene glycol, cottonseed oil and sesame oil.

The compositions of this invention can also be administered topically toa subject, i.e., by the direct laying on or spreading of the compositionon the epidermal or epithelial tissue of the subject. Such compositionsinclude, for example, lotions, creams, solutions, gels and solids. Thesetopical compositions may comprise a safe and effective amount, usuallyat least about 0.1%, or from about 1% to about 5%, of the4-oxoquinolizine compounds, and optionally also Polymyxins. Suitableexcipients for topical administration may optionally remain in place onthe skin as a continuous film, and resist being removed by perspirationor immersion in water. Generally, the excipient is organic in nature andcapable of having dispersed or dissolved therein the 4-oxoquinolizine.The excipient may include pharmaceutically-acceptable emolients,emulsifiers, thickening agents, and solvents and the like; these arewell known to the skilled artisan.

The 4-oxoquinolizine compounds and the pharmaceutical compositions ofthis invention can be administered topically or systemically. Systemicapplication includes any method of introducing the 4-oxoquinolizinecompounds into the tissues of the body, e.g. by intrathecal, epidural,intramuscular, transdermal, intravenous, intraperitoneal, subcutaneous,sublingual, rectal or oral administration. The specific dosage ofantimicrobial to be administered, as well as the duration of treatment,may be mutually dependent. The dosage and treatment regimen may alsodepend upon such factors as the specific 4-oxoquinolizine compound used,the resistance pattern of the infecting organism to the 4-oxoquinolizinecompound used, the ability of the 4-oxoquinolizine compound to reachminimum inhibitory concentrations at the site of the infection, thenature and extent of other infections (if any), the personal attributesof the subject (such as weight), compliance with the treatment regimen,the age and health status of the patient, and the presence and severityof any side effects of the treatment.

As an illustration, for a human adult (weighing approximately 70kilograms), from about 75 mg, or from about 200 mg, or from about 500 mgto about 30,000 mg, or to about 10,000 mg, or about 3,500 mg of a4-oxoquinolizine compound is administered per day. Treatment regimensmay extend from about 1 day to about 100 days, for example from about 3to about 56 days, such as from 3 to 20 days, in duration. Prophylacticregimens (such as avoidance of opportunistic infections inimmunocompromised patients) may extend 6 months, or longer, according togood medical practice.

One exemplary method of parenteral administration is through intravenousinjection. As is known and practiced in the art, all formulations forparenteral administration must be sterile. For mammals, especiallyhumans, (assuming an approximate body weight of 70 kilograms) individualdoses of from 100 to 3500 mg, for example from 500 mg to 7,000 mg, moreor to about 3,500 mg are typically acceptable.

In some cases, such as generalized, systemic infections or inimmune-compromised patients, the invention may be dosed intravenously.The dosage form is generally isotonic and at physiological pH. Thedosage amount will depend on the patient and severity of condition, aswell as other commonly considered parameters. Determination of suchdoses is well within the scope of practice for the skilled practitionerusing the guidance given in the specification.

An exemplary method of systemic administration is oral administration.Individual doses of from 20 to 500 mg, for example from 100 mg to 2,500mg may typically be useful

Topical administration can be used to deliver the 4-oxoquinolizinecompounds systemically, or to treat a local infection. The amounts of4-oxoquinolizine compounds to be topically administered may depend uponsuch factors as skin sensitivity, type and location of the tissue to betreated, the composition and excipient (if any) to be administered, theparticular 4-oxoquinolizine compound to be administered, as well as theparticular disorder to be treated and the extent to which systemic (asdistinguished from local) effects are desired.

The pharmaceutical composition may be in unit dosage form. In such formthe preparation is subdivided into unit doses containing appropriatequantities of the active component. The unit dosage form can be apackaged preparation, the package containing discrete quantities ofpreparation, such as packaged tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form. The composition can, if desired, also contain othercompatible therapeutic agents, discussed in more detail, below.

In therapeutic use, the 4-oxoquinolizine compounds or the compositionscomprising both Polymyxin and 4-oxoquinolizine compounds utilized in themethods of the invention may be administered to subjects at dosagelevels suitable to achieve therapeutic benefit. By therapeutic benefitis meant that the administration of compound leads to a beneficialeffect in the patient over time.

Initial dosages suitable for administration to humans may be determinedfrom in vitro assays or animal models. For example, an initial dosagemay be formulated to achieve a serum concentration that includes theIC₅₀ of the particular compound being administered, as measured in an invitro assay. Alternatively, an initial dosage for humans may be basedupon dosages found to be effective in animal models of the disease. Forexample, thr dose may vary depending on the symptoms, age, body weight,etc. of the patient. Usually, the 4-oxoquinolizine compounds areadministered to adults in a dose of 0.05 to 100 mg/kg/day, preferably0.1 to 50 mg/kg/day, in the systemic administration. When4-oxoquinolizine compounds are used for the local treatment, theconcentration of the active ingredient is 0.01 to 5%, preferably 0.1 to3%. Preferred dosages of Polymyxins are described herein elsewhere.

The 4-oxoquinolizine compounds and the Polymyxin may preferably beadministered simultaneously, however it is also contemplated within thescope of the present invention that the 4-oxoquinolizine compounds andthe Polymyxin is administrered sequentially in any order.

The Examples that follow are illustrative of specific embodiments of theinvention, and various uses thereof. They are set forth for explanatorypurposes only, and are not to be taken as limiting the invention.

EXAMPLES Example 1: Minimal Inhibitory Concentration (MIC) Determinationfor CDC Strains

This example shows that 4-oxoquinolizine compounds show potent activityagainst a pathogen that is a CDC category A or category B pathogen(Table 2) and for example B. thailandensis (Table 3).

The results for B. anthracis, F. tularensis, B. abortus, and B.pseudomallei are summarized in Table 2 below. The dilutions ranged from32 μg/ml to 0.015 μg/ml (12 2-fold dilution range). The final DMSOconcentration was 1.25% for B. anthracis and 2.5% for all other selectagents. The assay was performed in duplicate in 96-well plates with atotal assay volume of 100 μl. The bacteria were cultivated according tothe CLSI guidelines. The MIC value was determined as the lowestconcentration that resulted in no growth. The results for B.thailandensis are summarized in Table 3 below. A number of compoundsshowed a 2 to 4 dilutions improvement shift in MIC90 between pH7 andpH5. This property additionally makes the compounds therapeuticallyinteresting, particularly for use in acidic infected tissues and tissueswhere cytosolic acidity rises as a result of infection.

TABLE 2 showing potent antimicrobial activity against a panel of CDCpathogens exhibited by 4-oxoquinolizine compounds. MIC (ug/ml) B. F. B.B. Compounds Structures anthracis tularensis abortus pseudomallei 1

<0.008 <0.03  0.125 0.125 5

  0.125   0.125 1    2    6

  0.125   0.125 0.5  2    9

  0.06    0.125 0.25  0.5 

TABLE 3 showing the potent antimicrobial activity against B.thailandensis exhibited by 4- oxoquinolizine compounds and a 2 to 4dilutions improvement shift in MIC90 between pH7 and pH5. Dilutionsimprovement shift on B. MIC90 thailandensis B. thailandensis MIC90(between Compounds Structure (ug/ml) pH 7 and pH 5) 1

<0.048 Yes, 4× 2

<0.048 3

<0.048 5

0.390 6

0.390 No 7

<0.048 Yes, 4× 8

<0.024 10

<0.048 Yes, 2× 11

<0.048 12

0.390 13

0.195 14

<0.048 15

<0.048 16

<0.048

Example 2: Minimal Inhibitory Concentration (MIC) Determination forGram-Positive and Gram-Negative Bacteria

Example 2 shows the potent antibacterial activity of a number of4-oxoquinolizine compounds against a standard panel of Gram-positive andGram-negative strains (Table 4) in comparison to a well-known marketedquinolone (Ciprofloxacin) and glycopeptide (Vancomycin). Table 5 belowshows the resistant strains used for the data displayed in Table 6 belowthat shows the activity of a selection of 4-oxoquinolizine compoundsagainst a panel of quinolone-resistant strains.

MICs were determined following the standard CLSI protocol as describedherein below in Example 4 using doubling dilutions of compounds (0.03 to32 μg/ml), Levofloxacin, Ciprofloxacin, Vancomycin (0.125 to 128 μg/ml)in cation adjusted Mueller Hinton broth (CAMHB, Oxoid). The MIC wasdetermined as the lowest concentration of an individual drug that leadto no visible growth.

Table 4 below shows the MIC values for the instant 4-oxoquinolizinesagainst a standard panel of Gram-positive and Gram-negative bacteria incomparison to a well-known marketed quinolone (Ciprofloxacin) andglycopeptide (Vancomycin). Table 5 below shows the resistant strainsused for the data displayed in Table 6 below that shows the activity ofthe instant 4-oxoquinolizine compounds against a panel ofquinolone-resistant strains.

TABLE 4 showing the MIC values for a selection of 4-oxoquinolizinesagainst standard panels of Gram-positive and Gram -negative bacteria incomparison to a well-known marketed quinolone (Ciprofloxacin) andglycopeptide (Vancomycin). MIC (ug/ml) S. aureus S. aureus S. aureus S.epidermis E. faecalis E. faecium Compounds Structures 25923 25923 HS 10112228 29212 19434 Compound 2

<0.008 0, 125 0, 125 <0.008 <0.008 0, 5 Compound 3

<0.008 1 0, 06 0, 008 0, 016 0, 5 Compound 4

0, 06 0, 125 4 0, 06 0, 125 2 Compound 5

0, 03 0, 125 4 0, 06 0, 125 2 Compound 6

0, 03 0, 25 2 0, 03 0, 125 4 Compound 7

<0.008 1 1 <0.008 0, 06 2 Compound 8

<0.008 0, 25 0, 5 <0.008 0, 03 0, 5 Vancomycin Vancomycin 2 2 2 2 2 1Ciprofloxacin Ciprofloxacin 1 1 128 0, 25 1 8 MIC (ug/ml) E. cloacae P.aeruginosa A. baumanii Compounds Structures E. coli 25922 E. coli IMP13047 27853 19606 Compound 2

<0.03 <0.008 0, 25 2 0, 06 Compound 3

<0.03 0, 06 0, 5 2 0, 25 Compound 4

<0.03 <0.008 0, 06 0, 5 0, 25 Compound 5

<0.03 <0.008 0, 06 0, 5 0, 25 Compound 6

<0.03 0, 03 0, 125 2 0, 5 Compound 7

0, 03 <0.008 1 4 0, 5 Compound 8

0, 03 <0.008 0, 25 1 0, 125 Vancomycin Vancomycin >128 >128 >128 >128Ciprofloxacin Ciprofloxacin <0.125 <0.125 <0.125 0, 5 4

TABLE 5 listing the resistant strains used for the data displayed inTable 6 Organism Strain Resistance Phenotype A. baumannii OXA-51 CIP R,ESBL E. coli CTX-M-15 CIP R, ESBL E. coli 2906 CIP R Klebsiella sppKPC-3 CIP R, ESBL Pseudomonas spp PER-1 CIP R, ESBL P. aeruginosa1388-3-02 CIP R, POL R P. aeruginosa 143-3-03 CIP I, POL R S. aureusSMITH CIP R S. pneumoniae 49619 LEV S S. pneumonia 1027 LEV R CIP,ciprofloxacin; POL, Polyoxin; ESBL, extended spectrum beta-lactamases

TABLE 6 showing the activity of 4-o against a panel ofquinolone-resistant strains MIC (ug/ml) A. Baumanii E. coli E. coliKlebsiella S. aureus Compounds Structures OXA-51 CTX-M-15 2906 ssp KPC-3SMITH Compound 2

4 >32 32 8 <0.03 Compound 3

8 >32 >32 8 0, 5 Compound 4

16 16 8 4 0, 125 Compound 5

8 16 16 8 0, 125 Compound 6

8 16 16 16 0, 125 Compound 7

8 32 32 16 0, 25 Compound 8

4 >32 >32 16 <0.03 Ciprofloxacin Ciprofloxacin 128 128 128 128 32Levofloxacin Levofloxacin 16 16 32 >128 8 MIC (ug/ml) Pseudomonas P.aeruginosa P. aeruginosa S. pneumoniae S. pneumoniae CompoundsStructures ssp PER-1 1388-3-02 143-3-03 49619 1027 Compound 2

16 8 2 <0.03 0, 5 Compound 3

32 8 4 0, 5 2 Compound 4

8 16 4 0, 125 1 Compound 5

8 16 4 0, 125 2 Compound 6

16 32 8 0, 125 2 Compound 7

16 16 4 0, 25 4 Compound 8

16 8 2 <0.03 2 Ciprofloxacin Ciprofloxacin 16 16 2 0, 5 32 LevofloxacinLevofloxacin 32 32 8 0, 5 16

Example 3: Minimal Inhibitory Concentration (MIC) Determination forAcinetobacter Strains

Example 3 shows that 4-oxoquinolizine compounds show potent activityagainst a pathogen that is resistant against one or more of quinolones,carbapenems, aminosides and glycopeptides antibiotics, and specifically,The antibacterial activity of the compounds was assessed against avariety of A. bacter strains consisting mostly of resistant clinicalisolates (Table 7 and 8).

Bacterial Strains

Organisms used were one type strain and clinical isolates ofAcinetobacter baumannii from commercial culture collections or from K.Towner, Nottingham, UK. One Acinetobacter sp. (H064200250) was obtainedfrom D. Livermore, London, UK (see Table 7 below which lists theAcinetobacter strains referred to in Table 8 below).

Antimicrobial Agents

Ciprofloxacin, Levofloxacin, Vancomycin and Amikacin were purchased fromFluka and Aldrich (Sigma-Aldrich, Buchs, Switzerland); moxifloxacin,meropenem and imipenem/cilastatin were purchased from Apin ChemicalsLtd. (Abingdon, Oxon, UK).

Minimal Inhibitory Concentration (MIC) Determination

MICs were determined following the standard CLSI protocol as describedherein below in Example 4 using doubling dilutions of compounds (0.03 to32 μg/ml), Levofloxacin, Ciprofloxacin, Moxifloxacin, Amikacin,Vancomycin, Meropenem and Imipenem/Cilastatin (0.125 to 128 μg/ml) incation adjusted Mueller Hinton broth (CAMHB, Oxoid). Additionally, MICswere run in CAMHB supplemented. CLSI breakpoints were used to classifyresistance to carbapenems, quinolones, aminosides, and glycopeptides¹.The Acinetobacter strains were grown in CAMHB for 20-24 hours at 37° C.in ambient air. The MIC was determined as the lowest concentration of anindividual drug that lead to no visible growth. Referring to thefollowing tables, Table 7 shows the Acinetobacter strains used and Table8 below shows the activity of the present 4-oxoquinolizines againstdifferent Acinetobacter strains known to be resistant against a numberof well-known marketed antibiotic drugs such as quinolones, carbapenemsand aminoside antibiotics.

TABLE 7 showing the Acinetobacter strains used for the data displayed inTable 8 below. Resistance Organism Strain Source Origin Phenotype A.baumannii ATCC DSMZ¹ Reference — 19606 strain A. baumannii J2 Rocheunknown LEV R, CIP R Acinetobacter H064200250 D. Livermore² unknown LEVR, CIP R, MRP R, sp. (OXA51 IMP I upregulated) A. baumannii A329 K.Towner³ Barcelona, LEV R, CIP R, MRP R, ES IMP R, AMK R, TET R A.baumannii A387 K. Towner Ioannina, GR LEV I, CIP R, IPM R, AMK R, TET RA. baumannii A390 K. Towner Pleven, LEV R, CIP R, MRP R, Bulgaria IPM I,AMK R, TET I A. baumannii A401 K. Towner Taiwan LEV R, CIP R, AMK R, TETR A. baumannii A472 K. Towner Warsaw, PL LEV R, CIP R, MRP I, IMP I, AMKR, TET R A. baumannii A473 K. Towner Warsaw, PL LEV R, CIP R, MRP R, IMPR, AMK R, TET R A. baumannii A489 K. Towner Nottingham, LEV R, CIP R,MRP R, UK (patient IMP R, AMK R, TET R repatriated from Crete) LEV,Levofloxacin; CIP, Ciprofloxacin; MRP, Meropenem; IPM, Imipenem; AMK,Amikacin; TET, tetracycline ¹DSMZ, German Collection Strain ofMicroorganisms and Cell Cultures, Braunschweig, Germany. ²D. Livermore,Health Protection Agency, London, UK. ³K. Towner, Nottingham UniversityHospitals NHS Trust, Nottingham, UK.

TABLE 8 showing the MIC values for 4-oxoquinolizines against a panel ofAcinetobacter resistant strains in comparison to the efficacy of knowndrugs Levofloxacin, Moxifloxacin, Meropenem, Imipenem and Amikacin. MIC(ug/ml) A. baumannii A. baumannii Acinetobacter A. baumannii A.baumannii Compound Structure ATCC 19606 J2 spp. H064200250 A329 A489 1

0.25 8 8 16 4 2

≦0.03 4 4 8 2 3

0.25 8 8 16 4 4

0.25 8 16 16 8 5

0.25 8 16 16 4 6

0.5 16 8 16 8 7

0.5 16 8 16 8 8

1 8 2 16 2 Levofloxacin Levofloxacin 1 32 16 32 16 MoxifloxacinMoxifloxacin 0.5 32 16 32 16 Meropenem Meropenem 2 1 32 128 16 ImipenemImipenem 2 4 16 128 128 Amikacin Amikacin 16 32 16 >128 >128 MIC (ug/ml)A. baumannii A. baumannii A. baumannii A A. baumannii A. baumanniiCompound Structure A387 A390 401 A472 A473 1

1 16 4 8 4 2

0.5 8 4 4 8 3

1 >32 8 32 4 4

2 8 16 4 8 5

2 8 16 8 8 6

4 8 16 8 8 7

2 8 4 4 4 8

2 16 32 4 2 Levofloxacin Levofloxacin 8 16 16 16 16 MoxifloxacinMoxifloxacin 4 16 32 16 16 Meropenem Meropenem 4 32 32 8 16 ImipenemImipenem 16 16 32 16 32 Amikacin Amikacin >128 >128 >128 >128 >128

Example 4: Antimicrobial Activity of 4-Oxoquinolizines in Combinationwith Sub-Inhibitory Concentrations of Polymyxin B Against ClinicalIsolates Including Those which are Resistant to Quinolones, Carbapenemsand Other Antimicrobial Agents

Example 4 describes the determination of antimicrobial activity of4-oxoquinolizine compounds with 8-aniline and 8-aniline-likesubstitutions and shows that these exhibited potent activities inpresence of sub-inhibitory concentrations of polymyxin B against A.baumannii which are resistant to quinolones, carbapenems and otherantimicrobial agents. The inventive 4-oxoquinolizine compounds withaniline or aniline-like substitutions exhibited potent activities inpresence of sub-inhibitory concentrations of polymyxin B against A.baumannii including quinolone and multi-resistant strains. Theactivities of non-aniline amines 8-substitutions compounds and compound10 (ABT-719) were less potentiated by polymyxin B. The potentantimicrobial activity shown by the instant compounds when combined withsub-inhibitory concentrations of polymyxin B against Acinetobacterbaumannii including multi-resistant clinical isolates are shown in Table9, 10 and 11 below. Additionally, the 4-oxoquinolizine compounds exhibitbactericidal activity in absence or presence of polymyxin B asdetermined using a preliminary MBC assay. MIC ranges without and withsub-inhibitory concentrations of polymyxin B against resistant strainsof A. baumannii and one sensitive strain of Acinetobacter sp are shownin Table 12.

Bacterial Strains

Organisms used were one type strain and clinical isolates ofAcinetobacter baumannii from commercial culture collections or from K.Towner, Nottingham, UK. One Acinetobacter sp. (H064200250) was obtainedfrom D. Livermore, London, UK.

Antimicrobial Agents

The 4-oxoquinolizines with phenyl anilines and phenyl amines as8-substitutions as well as reference compound 10 (ABT-719) (see Table 9below) were synthesized. Polymyxin B sulfate, levofloxacin and amikacinwere purchased from Fluka (Sigma-Aldrich, Buchs, Switzerland);moxifloxacin, meropenem and imipenem/cilastatin were purchased from ApinChemicals Ltd. (Abingdon, Oxon, UK).

TABLE 9 Antimicrobial agents used and ranges tested Compounds StructureRange tested (μg/ml) Compound 1

0.03-32 Compound 17

0.03-32 Compound 2

0.03-32 Compound 3

0.03-32 Compound 7

0.03-32 Compound 8

0.03-32 Compound 4

0.03-32 Compound 5

0.03-32 Compound 6

0.03-32 Compound 10 (ABT-719)

0.008-8  Levofloxacin 0.125-128 Moxifloxacin 0.125-128 Meropenem0.125-128 Imipenem/cilastatin 0.125-128 Amikacin 0.125-128 Polymyxin B0.125-128

Minimal Inhibitory Concentration (MIC) Determination

MICs were determined following the standard CLSI protocol (Clinical andLaboratory Standards Institute. Methods for dilution antimicrobialsusceptibility tests for bacteria that grow aerobically. ApprovedStandard-seventh edition M7-A7, Clinical and Laboratory StandardsInstitute, Wayne, Pa., USA, 2006) using doubling dilutions of the8-phenyl anilines and 8-phenyl amines (0.03 to 32 μg/ml), referencecompound 10 (ABT-719) (0.008 to 8 μg/ml), and polymyxin B, levofloxacin,moxifloxacin, amikacin, meropenem and imipenem/cilastatin (0.125 to 128μg/ml) in cation adjusted Mueller Hinton broth (CAMHB, Oxoid).Additionally, MICs were run in CAMHB supplemented with sub-inhibitoryconcentrations (0.25×MIC) of polymyxin B sulfate. CLSI breakpoints wereused to classify resistance to carbapenems, quinolones, aminoglycosides,and tetracyclines (DSMZ, German Collection Strain of Microorganisms andCell Cultures, Braunschweig, Germany). The Acinetobacter strains weregrown in CAMHB for 20-24 hours at 37° C. in ambient air. The MIC wasdetermined as the lowest concentration of an individual drug that leadto no visible growth.

Since trailing was observed in the first experiment, MICs were confirmedby adding 10 μl alamarBlue (alamarBlue™ Assay, Biosource; LucernaChemAG) to each well in the 2nd experiment. The alamarBlue Assayincorporates a growth indicator based on detection of metabolicactivity. Reduction related to growth causes the redox indicator tochange from oxidized (blue) form to reduced (red) form. Followingincubation at 37° C. for 1 hour, the MIC was read as lowestconcentration of an individual drug that lead to no growth indicated byblue color.

Preliminary Minimal Bactericidal Concentration (MBC) Determination

After reading the MICs, microtiter plates were shaken (700 rpm, 5minutes) and 5 μl culture from each well from the plates used for theMIC determination was spotted on Mueller-Hinton agar plates withoutantibiotic. The plates were incubated for 18-20 hours at 37° C. The MBCwas read as the lowest concentration at which colony growth was ca. 90%less than the positive control (i.e. usually where single colonies or nocolonies were observed).

Mics

MIC data are shown in Table 10 (1st experiment) and Table 11 (2ndexperiment) and MIC ranges are summarized in Table 12 below. All A.baumannii strains except for the type strain ATCC 19606 and oneAcinetobacter sp. were resistant to ciprofloxacin and levofloxacin (onestrain, A. baumannii A387 exhibited intermediate susceptibility againstlevofloxacin and resistance to ciprofloxacin). Additionally, most of thestrains were resistant to carbapenems, aminoglycosides, andtetracycline. MICs for polymyxin B were ranging from 0.25 to 2 μg/ml(1st experiment) and 0.25 to 0.5 μg/ml (2nd experiment).

TABLE 10 MICs and preliminary MBCs without and with sub-inhibitoryconcentrations of polymyxin B (PB) against 11 strains of A. baumanniiand one strain of Acinetobacter sp. (1^(st) experiment). Chemical namesand structures of the compounds used are given in Table 1 herein above.A. baumannii ATCC 19606 without PB +0.06 μg/ml PB MIC without CompoundsMIC MBC MIC MBC PB/MIC + PB Compound 1 0.25 0.25 ≦0.03 ≦0.03 ≧8 Compound17 0.5 1 ≦0.03 ≦0.03 ≧16 Compound 2 ≦0.03 ≦0.03 ≦0.03 ≦0.03 ≧1 Compound3 0.25 0.125 ≦0.03 ≦0.03 ≧8 Compound 7 0.5 0.5 ≦0.03 ≦0.03 ≧16 Compound8 1 0.125 ≦0.03 ≦0.03 ≧32 Compound 4 0.25 0.125 0.125 0.06 2 Compound 50.25 0.125 0.06 0.06 4 Compound 6 0.5 0.25 0.06 0.06 8 Compound 10(ABT-719) ND ND 0.03 0.03 256 Levofloxacin 1 0.5 1 1 1 Moxifloxacin 0.50.5 0.5 0.5 1 Meropenem 2 128 1 2 2 Imipenem 2 2 1 2 2 Amikacin 16 16 88 2 Polymyxin B 0.5 0.5 0.5 0.5 1 A. baumannii J2 without PB +0.125μg/ml PB MIC without Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 88 ≦0.03 ≦0.03 ≧256 Compound 17 4 4 0.06 0.06 64 Compound 2 4 4 ≦0.03≦0.03 ≧128 Compound 3 8 8 0.125 0.125 64 Compound 7 16 2 ≦0.03 ≦0.03≧512 Compound 8 8 8 0.125 0.125 64 Compound 4 8 4 2 1 4 Compound 5 8 80.5 0.5 16 Compound 6 16 8 1 1 16 Compound 10 (ABT-719) 2 1 1 1 2Levofloxacin 32 32 8 8 4 Moxifloxacin 32 32 8 8 4 Meropenem 1 2 1 1 1Imipenem 4 4 ≦0.125 ≦0.125 ≧32 Amikacin 32 32 4 4 8 Polymyxin B 0.5 0.50.5 0.5 1 A. baumannii NCTC 13301 without PB +0.125 μg/ml PB MIC withoutCompounds MIC MBC MIC MBC PB/MIC + PB Compound 1 32 32 ≦0.03 0.06 ≧1024Compound 17 16 16 ≦0.03 ≦0.03 ≧512 Compound 2 16 8 ≦0.03 ≦0.03 ≧512Compound 3 >32 >32 0.5 0.5 ≧64 Compound 7 16 16 0.06 0.06 256 Compound 832 16 0.5 1 64 Compound 4 8 8 1 0.5 8 Compound 5 16 8 1 1 16 Compound 632 16 1 4 (1)* 32 Compound 10 (ABT-719) 8 2 1.00 1.00 8 Levofloxacin 3232 16 16 2 Moxifloxacin 128 32 128 128 1 Meropenem 32 32 1 1 32 Imipenem128 128 16 16 8 Amikacin >128 >128 >128 >128 ≧1 Polymyxin B 0.5 0.5 10.5 0.5 Acinetobacter spp. H064200250 without PB +0.125 μg/ml PB MICwithout Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 8 8 ≦0.03 0.25(0.06)* ≧256 Compound 17 8 8 0.06 0.06 128 Compound 2 4 4 ≦0.03 ≦0.03≧128 Compound 3 8 8 0.5 0.5 16 Compound 7 8 4 0.25 0.25 32 Compound 8 24 0.06  0.5 (0.125)* 32 Compound 4 16 16 4 8 4 Compound 5 16 8 1 1 16Compound 6 8 8 4 4 2 Compound 10 (ABT-719) 2 0.5 1 1 2 Levofloxacin 1616 16 16 1 Moxifloxacin 16 16 16 16 1 Meropenem 32 16 1 1 32 Imipenem 1616 0.5   2 (0.5)* 32 Amikacin 16 16 16 64 (16)* 1 Polymyxin B 0.5 0.50.25 0.5 2 A. baumannii A14 without PB +0.06 μg/ml PB MIC withoutCompounds MIC MBC MIC MBC PB/MIC + PB Compound 1 32 32 0.06 0.06 512Compound 17 16 16 1 1 16 Compound 2 16 16 ≦0.03 ≦0.03 ≧512 Compound3 >32 >32 1 1 ≧32 Compound 7 32 16 1 1 32 Compound 8 32 16 1 1 32Compound 4 32 16 8 8 4 Compound 5 32 16 4 8 8 Compound 6 32 16 8 8 4Compound 10 (ABT-719) 8 2 8.00 2.00 1 Levofloxacin 32 32 32 32 1Moxifloxacin 32 32 32 32 1 Meropenem 1 2 0.5 0.5 2 Imipenem 1 2 1 1 1Amikacin 32 32 16 16 2 Polymyxin B 0.5 0.5 0.5 0.5 1 A. baumannii A329without PB +0.125 μg/ml PB MIC without Compounds MIC MBC MIC MBCPB/MIC + PB Compound 1 16 16 NR NR Compound 17 8 8 NR NR Compound 2 8 8NR NR Compound 3 16 16 NR NR Compound 7 16 16 NR NR Compound 8 16 16 NRNR Compound 4 16 16 NR NR Compound 5 16 16 NR NR Compound 16 16 NR NRCompound 10 (ABT-719) 8 2 NR NR Levofloxacin 32 32 NR NR Moxifloxacin 3264 NR NR Meropenem 128 128 NR NR Imipenem 128 128 NR NRAmikacin >128 >128 NR NR Polymyxin B 0.5 0.5 NR NR A. baumannii A387without PB +0.06 μg/ml PB MIC without Compounds MIC MBC MIC MBC PB/MIC +PB Compound 1 1 1 ≦0.03 ≦0.03 ≧32 Compound 17 1 2 ≦0.03 ≦0.03 ≧32Compound 2 0.5 0.5 ≦0.03 ≦0.03 ≧16 Compound 3 1 1 ≦0.03  0.25 (≦0.03)*≧32 Compound 7 2 2 ≦0.03 ≦0.03 ≧64 Compound 8 2 2 ≦0.03 ≦0.03 ≧64Compound 4 2 2 1 1 2 Compound 5 2 1 0.25 0.25 8 Compound 6 4 4 0.5 0.5 8Compound 10 (ABT-719) 0.5 0.5 0.5 0.25 1 Levofloxacin 8 8 8 8 1Moxifloxacin 4 4 4 4 1 Meropenem 4 4 1 1 4 Imipenem 16 16 8 8 2Amikacin >128 >128 >128 >128 ≧1 Polymyxin B 0.25 0.25 0.25 0.25 1 A.baumannii A390 without PB +0.125 μg/ml PB MIC without Compounds MIC MBCMIC MBC PB/MIC + PB Compound 1 16 16 NR NR Compound 17 16 16 NR NRCompound 2 8 8 NR NR Compound 3 >32 >32 NR NR Compound 7 8 4 NR NRCompound 8 16 8 NR NR Compound 4 8 8 NR NR Compound 5 8 8 NR NR Compound6 8 8 NR NR Compound 10 (ABT-719) 1 1 NR NR Levofloxacin 16 8 NR NRMoxifloxacin 16 16 NR NR Meropenem 32 32 NR NR Imipenem 16 16 NR NRAmikacin >128 >128 NR NR Polymyxin B 0.25 0.25 NR NR A baumannii A 401without PB +0.06 μg/ml PB MIC without Compounds MIC MBC MIC MBC PB/MIC +PB Compound 1 4 16 (4)* ≦0.03 ≦0.03 ≧128 Compound 17 4 8 0.5 0.5 8Compound 2 4 2 ≦0.03 ≦0.03 ≧128 Compound 3 8 4 0.125 0.125 64 Compound 74 4 0.5 0.5 8 Compound 8 32 8 ≦0.03 ≦0.03 ≧1024 Compound 4 16 4 4 2 4Compound 5 16 8 2 2 8 Compound 6 16 4 4 4 4 Compound 10 (ABT-719) 2 11.00 1.00 2 Levofloxacin 16 16 16 16 1 Moxifloxacin 32 16 16 8 2Meropenem 32 16 4 4 8 Imipenem 32 32 8 8 4 Amikacin >128 >128 >128 >128≧1 Polymyxin B 1 1 0.5 0.5 2 A. baumannii A472 without PB +0.125 μg/mlPB MIC without Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 8 8≦0.03 ≦0.03 ≧256 Compound 17 4 4 ≦0.03 0.125 ≧128 Compound 2 4 4 ≦0.03≦0.03 ≧128 Compound 3 32 >32 0.5 0.25 64 Compound 7 4 4 ≦0.03 ≦0.03 ≧128Compound 8 4 2 ≦0.03 ≦0.03 ≧128 Compound 4 4 2 1 1 4 Compound 5 8 4 1 18 Compound 6 8 4 0.5 0.5 16 Compound 10 (ABT-719) 1 1 0.5 0.25 2Levofloxacin 16 8 4 4 4 Moxifloxacin 16 8 32 4 0.5 Meropenem 8 8 ≦0.125≦0.125 ≧64 Imipenem 16 16 0.25 0.25 64 Amikacin >128 >128 16 32 ≧8Polymyxin B 0.5 0.5 ≦0.125 ≦0.125 ≧4 A. baumannii A473 without PB +0.125μg/ml PB MIC without Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 44 NR NR Compound 17 4 4 NR NR Compound 2 8 2 NR NR Compound 3 4 4 NR NRCompound 7 4 4 NR NR Compound 8 2 2 NR NR Compound 4 8 4 NR NR Compound5 8 4 NR NR Compound 6 8 4 NR NR Compound 10 (ABT-719) 1 0.5 NR NRLevofloxacin 16 16 NR NR Moxifloxacin 16 8 NR NR Meropenem 16 64 (16)*NR NR Imipenem 32 32 NR NR Amikacin >128 >128 NR NR Polymyxin B 1 1 NRNR A. baumannii A489 without PB +0.25 μg/ml PB MIC without Compounds MICMBC MIC MBC PB/MIC + PB Compound 1 4 8 ≦0.03 ≦0.03 ≧128 Compound 17 8 4≦0.03 ≦0.03 ≧256 Compound 2 2 2 ≦0.03 ≦0.03 ≧64 Compound 3 4 2 0.06 0.0664 Compound 7 8 4 ≦0.03 ≦0.03 ≧256 Compound 8 2 2 ≦0.03 ≦0.03 ≧64Compound 4 8 4 1 1 8 Compound 5 4 2 0.5 1 8 Compound 6 8 8 0.5 1 16Compound 10 (ABT-719) 1 1 1.00 1.00 1 Levofloxacin 16 32 8 8 2Moxifloxacin 16 8 4 4 4 Meropenem 16 8 0.5 0.5 32 Imipenem 128 64 2 2 64Amikacin >128 >128 128 128 ≧1 Polymyxin B 2 2 1 1 2 PB, polymyxin B*skipped growth on MBC plate ND, MIC value was higher than previousdata, thus this value was not considered NR, MIC was not readable due toinhomogeneous growth in the wells (i.e. skipped growth in many wells)

TABLE 11 MICs and preliminary MBCs without and with sub-inhibitoryconcentrations of polymyxin B (PB) against 11 strains of A. baumanniiand one strain of Acinetobacter sp. (2^(nd) experiment). Chemical namesand structures of the compounds used are given in Table 1 herein above.A. baumannii ATCC 19606 without PB +0.06 μg/ml PB MIC without CompoundsMIC MBC MIC MBC PB/MIC + PB Compound 1 0.25 0.25 ≦0.03 ≦0.03 ≧8 Compound2 0.125 *(0.06) 0.125 ≦0.03 ≦0.03 ≧4 Compound 3 0.25 0.5 ≦0.03 ≦0.03 ≧8Compound 7 0.25 0.25 ≦0.03 ≦0.03 ≧8 Compound 8  0.25 *(0.125) 0.25 ≦0.03≦0.03 ≧8 Compound 4 0.5 0.5 ≦0.03 (0.06)^(#) ≦0.03 ≧16 Compound 5  0.5*(0.25) 0.25 0.06 0.06 8 Compound 6 0.25 0.25 0.06 0.06 4 Compound 10(ABT-719) 0.125 *(0.06) 0.125 0.06 0.03 2 Levofloxacin 1 1      1(0.5)^(#) 1.00 1 Moxifloxacin 1 1 0.5 0.5 2 Meropenem  1 *(64) 4    1(2)^(#) 2 1 Imipenem  1 *(4) 4 1 1 1 Amikacin 16 16 8 8 2 Polymyxin B0.5 0.5 0.25 0.25 2 A. baumannii J2 without PB +0.125 μg/ml PB MICwithout Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 16 16 ≦0.03≦0.03 ≧512 Compound 2 4 8 ≦0.03 ≦0.03 ≧128 Compound 3 32 32 ≦0.03 ≦0.03≧1024 Compound 7 4 4 ≦0.03 ≦0.03 ≧128 Compound 8 4 4 ≦0.03 ≦0.03 ≧128Compound 4 8 16 0.06 0.06 128 Compound 5 4 4 0.5 0.5 8 Compound 6 8 160.25 0.25 32 Compound 10 (ABT-719) 2 2 0.25 0.25 8 Levofloxacin 32 32 44 8 Moxifloxacin 32 32 8 8 4 Meropenem 2 2 ≦0.125 ≦0.125 ≧16 Imipenem 8(4)^(#) 8 ≦0.125 ≦0.125 ≧64 Amikacin 8 8 0.5 0.5 16 Polymyxin B 0.5 0.5≦0.125 ≦0.125 ≧4 A. baumannii NCTC 13301 without PB +0.125 μg/ml PB MICwithout Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 32 32 ≦0.03≦0.03 ≧1024 Compound 2 16 32 ≦0.03 ≦0.03 ≧512 Compound 3 >32 >32 ≦0.03≦0.03 ≧1024 Compound 7 16 16 ≦0.03 ≦0.03 ≧512 Compound 8 16 (8)^(#)  160.5 0.5 32 Compound 4 32 32 0.25 0.25 128 Compound 5 8 8 1 1 8 Compound6 16 8 1 2 16 Compound 10 (ABT-719) 2 2 0.5 0.5 4 Levofloxacin 32 168.00 4.00 4 Moxifloxacin 64 (32)^(#) 64 8 8 8 Meropenem 64 64 0.25≦0.125 256 Imipenem 128 128 1 0.5 128 Amikacin >128 >128 128 64 ≧1Polymyxin B 0.25 0.25 ≦0.125 *(0.25) ≦0.125 ≧2 Acinetobacter spp.H064200250 (OXA51) without PB +0.125 μg/ml PB MIC without Compounds MICMBC MIC MBC PB/MIC + PB Compound 1 16 32 ≦0.03 ≦0.03 ≧512 Compound 2 4 8≦0.03 (0.06)^(#) 0.06 ≧128 Compound 3 16 16 0.06 0.06 256 Compound 7 16(8)^(#)  8 ≦0.03 ≦0.03 ≧512 Compound 8 4 (8)# 4 0.125 0.125 32 Compound4 32 (16)^(#) 32 0.5 0.5 64 Compound 5 16 16 4 4 4 Compound 6 16 >32 2 18 Compound 10 (ABT-719) 1 1 1 0.5 1 Levofloxacin 16 16 8 8 2Moxifloxacin 16 16 16 16 1 Meropenem 16 16 0.25 0.25 64 Imipenem 8 8 0.50.5 16 Amikacin 8 8 1 1 8 Polymyxin B 0.25 0.25 0.25 0.25 1 A. baumanniiA14 without PB +0.06 μg/ml PB MIC without Compounds MIC MBC MIC MBCPB/MIC + PB Compound 1 32 32 0.06 0.06 512 Compound 2 16 16 0.06 0.06256 Compound 3 >32 >32 0.125 0.125 ≧256 Compound 7 16 16 2 2 8 Compound8 8 16 0.5 0.5 16 Compound 4 32 >32 2 8 16 Compound 5 16 16 4 4 4Compound 6 16 16 4 8 4 Compound 10 (ABT-719) 2 2 2 1 1 Levofloxacin 3232 16.00 16.00 2 Moxifloxacin 32 32 16 16 2 Meropenem 1 (2)^(#) 2 1 1 1Imipenem 1 1 1 0.5 1 Amikacin 16 16 8 8 2 Polymyxin B 0.25 0.25 0.250.25 1 A. baumannii A329 without PB +0.06 μg/ml PB MIC without CompoundsMIC MBC MIC MBC PB/MIC + PB Compound 1 16 32 2 2 8 Compound 2 8 8 0.5(1)^(#)   0.5 16 Compound 3  16 (>32)^(#) 32 0.5 (1)^(#)   0.25 32Compound 7 16 16 2 2 8 Compound 8 8 8 2 2 4 Compound 4 16 (32)^(#) 32 88 2 Compound 5  8 (16)^(#) 16 8 8 1 Compound 6 16 16 8 8 2 Compound 10(ABT-719) 2 2 1 (2)^(#) 2 2 Levofloxacin 16 32 32 16 0.5 Moxifloxacin 3232 32 32 1 Meropenem  128 (>128)^(#) >128 32 (64)^(#) 32 4Imipenem >128 >128 128 128 ≧1 Amikacin  128 (>128)^(#) 128 128 128 1Polymyxin B 0.25 0.25 0.25 0.25 1 A. baumannii A387 without PB +0.06μg/ml PB MIC without Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 44 ≦0.03 ≦0.03 ≧128 Compound 2 1 1 ≦0.03 ≦0.03 ≧32 Compound 3 1 4 ≦0.03≦0.03 ≧32 Compound 7 2 2 ≦0.03 ≦0.03 ≧64 Compound 8 1 1 0.5 0.25 2Compound 4 2 2  0.25 (0.125)^(#) 0.5 8 Compound 5 4 4 1 1 4 Compound 6 416 1 1 4 Compound 10 (ABT-719) 0.5 0.5 0.5 0.5 1 Levofloxacin 4 8 4.004.00 1 Moxifloxacin 8 8 8 8 1 Meropenem 8 8 4 (2)^(#) 2 2 Imipenem 16 168 8 2 Amikacin >128 >128 128 128 ≧1 Polymyxin B 0.25 0.25 ≦0.125(0.25)^(#)   0.25 ≧2 A. baumannii A390 without PB +0.06 μg/ml PB MICwithout Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 16 16 ≦0.03≦0.03 ≧512 Compound 2 16 16 ≦0.03 (0.06)^(#) 0.06 ≧512 Compound3 >32 >32 0.125 0.125 ≧256 Compound 7 16 16 0.125 0.125 128 Compound 816 16 0.5 0.5 32 Compound 4 32 32 0.5 0.5 64 Compound 5  8 (16)^(#) 16 22 4 Compound 6 16 (8)^(#) 8 2 2 8 Compound 10 (ABT-719) 1 1 0.5 0.5 2Levofloxacin 16 (8)^(#) 8 8 8 2 Moxifloxacin 16 16 8 8 2 Meropenem 32 322 2 16 Imipenem 16 16 4 4 4 Amikacin >128 >128 64 64 ≧2 Polymyxin B 0.250.25 ≦0.125 ≦0.125 ≧2 A baumannii A 401 without PB +0.06 μg/ml PBCompounds MIC MBC MIC MBC MIC without Compound 1 16 8 ≦0.03 ≦0.03 ≧512Compound 2 4 4 ≦0.03 ≦0.03 ≧128 Compound 3 16 16 0.125 0.125 128Compound 7 8 8 1 0.5 8 Compound 8 8 8 0.25 0.25 32 Compound 4 16 32 0.50.5 32 Compound 5 8 8 4 4 2 Compound 6 16 16 4 32 4 Compound 10(ABT-719) 1 1 1 1 1 Levofloxacin 16 16 16.00 8.00 1 Moxifloxacin 16 1616 8 1 Meropenem 16 32 4 4 4 Imipenem 32 32 4 4 8Amikacin >128 >128 >128 >128 ≧1 Polymyxin B 0.5 0.5 0.25 0.25 2 A.baumannii A472 without PB +0.125 μg/ml PB Compounds MIC MBC MIC MBC MICwithout Compound 1 8 32 ≦0.03 ≦0.03 ≧256 Compound 2 4 8 ≦0.03 ≦0.03 ≧128Compound 3 >32 >32 ≦0.03 ≦0.03 ≧1024 Compound 7 8 8 ≦0.03 ≦0.03 ≧256Compound 8 4 4 0.06 0.06 64 Compound 4 16 16 0.125 0.125 128 Compound 54 4 1 1 4 Compound 6 4 8 1 1 4 Compound 10 (ABT-719) 1 1 0.5 0.5 2Levofloxacin 8 8 4 4 2 Moxifloxacin 16 16 4 4 4 Meropenem 8 8 ≦0.125≦0.125 ≧64 Imipenem 8 8 0.25 0.25 32 Amikacin >128 >128 32 32 ≧4Polymyxin B 0.5 0.5 0.25 0.25 2 A. baumannii A473 without PB +0.06 μg/mlPB MIC without Compounds MIC MBC MIC MBC PB/MIC + PB Compound 1 8 8 1 18 Compound 2 4 4   0.06 (0.125)^(#) 0.125 64 Compound 3 8 8 ≦0.03(0.06)^(#) 0.06 ≧256 Compound 7 4 4 0.06 0.06 64 Compound 8 4 4 0.5 0.58 Compound 4 8 8 0.25 0.25 32 Compound 5 8 8 1 1 8 Compound 6 8 8 2 2 4Compound 10 (ABT-719) 0 1 0.5 0.5 0 Levofloxacin  8 (16)^(#) 8 8.00 8.001 Moxifloxacin 8 8 8 8 1 Meropenem 16 (32)^(#) 16 2 2 8 Imipenem 32 32 44 8 Amikacin >128 >128 128 128 ≧1 Polymyxin B 0.25 (2)^(#)   0.25 0.250.25 1 A. baumannii A489 without PB +0.25 μg/ml PB MIC without CompoundsMIC MBC MIC MBC PB/MIC + PB Compound 1 16 16 ≦0.03 ≦0.03 ≧512 Compound 24 4 ≦0.03 ≦0.03 ≧128 Compound 3 4 4 ≦0.03 ≦0.03 ≧128 Compound 7 8 16≦0.03 ≦0.03 ≧256 Compound 8 8 8 ≦0.03 ≦0.03 ≧256 Compound 4 16 16 ≦0.03(0.06)^(#) 0.06 ≧512 Compound 5 8 (4)^(#) 8 0.5 0.5 16 Compound 6 4 40.5 0.5 8 Compound 10 (ABT-719) 1 1 0.5 1 2 Levofloxacin 32 (16)^(#) 168 4 4 Moxifloxacin 16 16 4 4 4 Meropenem 16 (32)^(#) 16 ≦0.125 ≦0.125≧128 Imipenem 64 64 0.25 0.25 256 Amikacin >128 >128 64 64 ≧2 PolymyxinB 0.5 0.5 0.25 0.25 2 PB, polymyxin B ^(#)MIC read with alamar blue

TABLE 12 MIC ranges without and with sub-inhibitory concentrations ofpolymyxin B against resistant strains of A. baumannii and one strain ofAcinetobacter sp.¹ MIC range MIC range with Range ratios MIC withoutwithout PB PB (0.25 × MIC) PB/MIC with PB 01-10-2008 14-10-200801-10-2008 14-10-2008 01-10-2008 14-10-2008 Compounds Structure n = 8² n= 11³ n = 8 n = 11 n = 8 n = 11 Compound 1

  1-32   4-32 ≦0.03- 0.06 ≦0.03- 2 ≧32- ≧1024   8-≧1024 Compound 17

  1-16 ND⁴ ≦0.03- 1 ND 8- ≧512 ND Compound 2

0.5-16   1-16 ≦0.03- 1 ≦0.03- 0.5 ≧16- ≧512  16-≧512 Compound 3

  1-≧32   1-≧32 ≦0.03- 1 ≦0.03- 0.5 16- ≧64  32-≧1024 Compound 7

  2-32   2-16 ≦0.03- 1 ≦0.03- 2 8- ≧512   8-≧512 Compound 8

  2-32   2-32 ≦0.03- 1 ≦0.03- 8 32- ≧1024   2-≧512 Compound 4

  2-32   4-16 1- 8 0.5- 8 2- 8   1-16 Compound 5

  2-32   4-16 0.25- 4 0.25- 8 8- 16   4-32 Compound 6

  4-32 ND 0.5- 8 ND 2- 32 ND Compound 10 (ABT-719)

0.5-8 0.5-2 0.5- 8 0.25- 2 1- 8   1-8 Levofloxacin   8-32   4-32 4- 4-1- 0.5-8 32 32 4 Moxifloxacin   4-128   8-64 4- 4- 0.5-   1-8 128 32 4Meropenem   1-32   1-128 <0.125- ≦0.125- 1-   1-256 4 32 ≧64 Imipenem/  1-128   1->128 <0.125- ≦0.125- 1-   1-256 cilastatin 16 128 64Amikacin  16->128   8->128 4- 0.5- 1-   1-16 >128 >128 ≧8 ¹ A. baumanniiATCC 19606 was not taken for the calculation of MIC ranges since it wasvery susceptible against the 4-oxoquinolizine compounds. ²n = 8, threestrains (A. baumannii A329, A390 and A473) were not taken for thecalculation of MIC ranges since no homogenous growth (many skippedwells) was observed in presence of polymyxin B. ³n = 11, all strainsexcept for the ATCC strain were taken for the calculation of MIC ranges⁴ND, not determined

The 8-phenyl anilines compounds 1-8 and reference compound 10 (ABT-719)exhibited potent activities against the A. baumanii ATCC 19606 strainwith and without sub-inhibitory concentrations of polymyxin B. Sincethis strain was more susceptible compared to the clinical isolates, itsMIC values were not taken into account for calculations for MIC ranges(Table 12).

Reference compound 10 (ABT-719) was more active against the quinoloneresistant strains (MIC range: 0.5 to 8 μg/ml) than the 8-phenyl anilines(MIC ranges: 1 to >32 μg/ml) in the absence of polymyxin B.

Three A. baumannii strains did not grow homogenously in all wells of themicrotiter plates containing 0.125 μg/ml polymyxin B in the firstexperiment (A. baumannii A329, A390, A473), thus these strains were notconsidered for the calculation of MIC ranges in Table 12. In the 2ndexperiment, a lower polymyxin B concentration was used (0.06 μg/ml) andthe growth was homogenous in all wells.

The activity of the 8-phenyl anilines compounds 1, 2, 3, 7, 8 and 17 waspotentiated in the presence of polymyxin B. The compounds exhibitedlower MICs ranging from 50.03 to 8 μg/ml in the presence ofsub-inhibitory concentrations of polymyxin B compared to compound 10(ABT-719) and the 8-phenyl amines (compounds 4, 5, and 6) for which MICswere ranging from 0.25 to 8 μg/ml. Thus, the activities of the 8-phenylanilines were 2- to >1000-fold greater in presence of polymyxin Bcompared to activities without polymyxin B (Table 12). Overall, MICs ofthe 8-phenyl anilines were 50.03 to 0.5 μg/ml for the majority ofstrains except for 4-fold greater MICs against A. baumannii A-329 (MICswere 0.5 to 2 μg/ml). For this strain, MICs of the 8-phenyl anilineswere less affected by the presence of polymyxin B (2- to 32-fold lowerMICs in presence of polymyxin B) in contrast to the other strains (Table11). Compound 2 and 3 were the most active 8-phenyl anilines in presenceof polymyxin B against the quinolone resistant strains of A. baumanniiwith MICs ranging from 50.03 to 0.5 μg/ml (Table 12). However, compound2 was more active against the quinolone resistant Acinetobacter strainsthan compound 3 in absence of polymyxin B (Table 10 and 11).

The activities of levofloxacin, moxifloxacin or amikacin were notsignificantly affected by polymyxin B, whereas MICs of meropenem andimipenem/cilastatin were 1- to 256-fold lower in presence of polymyxin B(Table 12).

Preliminary MBCs

MBCs were estimated by spotting 5 μl of cultures from the MIC plates onagar. Preliminary MBC data are shown in Table 10 (1st experiment) andTable 11 (2nd experiment). The 8-phenyl anilines, 8-phenyl amines andthe reference compound 10 exhibited apparent bactericidal activitiesagainst A. baumannii with preliminary MBC/MIC ratios ranging from 1 to 4except for a few compounds vs. a few strains with a MBC/MIC ratio of 8(Table 10 and 11). There was no difference observed in MBC/MIC ratios inabsence or presence of polymyxin B, although the absolute MBC (and MIC)values were lower in presence of polymyxin B especially for the 8-phenylanilines.

Example 5: Antimicrobial Activity of 4-Oxoquinolizines Against aSelected Panel of Gram-Positive and Gram-Negative Strains and K.pneumonia

Example 5 shows the antibacterial activity of particular4-oxoquinolizines against a selection of Gram-positive and Gram-negativestrains (Table 13) and of a selection of three 4-oxoquinolizines on K.pneumonia strains (Table 14).

MIC Values for all compounds were measured against a selection ofbacterial strains. The MICs of 4-oxoquinolizines against a selection ofGram-negative and Gram-positive bacterial strains are shown in Table 13in comparison with known antibiotics. Table 14 shows the activity of aselected number of 4-oxoquinolizines against strains of K. pneumonia,one sensitive and one quinolone-resistant strain (NDM-1 BAA-2146).

TABLE 13 showing the MIC value of some 4-oxoquinolizines on a selectedpanel of bacterial strains. Chemical names and structures of thecompounds are given in Table 1 herein above. MICs (μg/mL) against Grampositive bacteria MICs (μg/mL) S. aureus against Gram negative bacteriaS. aureus BAA-1556 B. thailandensis E. coli P. aeruginosa A. baumanniiS. aureus ATCC 43300 (USA300-Type Compounds E264 ATCC 25922 ATCC 27853ATCC 19606 ATCC 25923 (MRSA, FQ-S) MRSA, FQ-R) Compound 4 1 0,016 1 0,250,064 0.032-0.064 0,5 Compound 13K salt 0,5 0,016 1 0, 125 0,016 0,0160,5 Compound 54 4 0,25 >8 4 0.064-0.125 0,064 0,5 Compound 6K salt 40.064-0.125 4-8 1 0, 125 0,064 1 Compound 55K salt 8 0,5 >8 8 0,50.125-0.25 8 Compound 56K salt 0,25 0,032 2 0, 125 0,008 0,008 0, 125Compound 57 >8 1 >8 8 0.25-0.5 0,25 8 Compound 58K salt 1 0,25 >8 10,064 0,064 1-2 Compound 59K salt 0,5 0,064 4 0,25 0,032 0,016 0,5Compound 60K salt 0,5 0,032 2 0,064 0,008 ≦0.004 0,125 Compound 61K salt0,5 0,032 2 0, 125 ≦0.004 ≦0.004 0,064 Compound 62K salt 2 0,032 1 10,25 0, 125 4 Compound 63 2 0,5 >8 2 0,064 0,032 1 Compound 64K salt 20, 125 8 0,5 0,032 0,032 0,5 Compound 65K salt 0,5 0.032-0.064 4 0,250,016 0,008 0,25 Compound 66K salt 1 0, 125 4 0,5 0,008 0,008 0,25Compound 67K salt 0,25 0,032 2 0,25 0,016 0,016 1 Compound 68K salt 10,064 2 0,25 ≦0.004 ≦0.004 0,25 Compound 69K salt 1 0, 125 4 0.25-0.5≦0.004 ≦0.004 0,25 Compound 70K salt 0,25 0,032 1 0, 125 ≦0.004 ≦0.0040,064 Compound 71K salt 2 0, 125 >8 0,5 ≦0.004 ≦0.004 0, 125 Compound72K salt 2 0.008-0.016 0,5 0,25 0,008 0.008-0.016 0,25 Compound 73K salt0,5 0,032 2 0, 125 ≦0.004 ≦0.004 0,064 Compound 74K salt 2 0.064-0.125 80,5 0,016 0,016 0,5 Compound 75K salt 0,25 0,032 1 0,064 ≦0.004 ≦0.0040,032 Compound 76K salt 2 0,032 1 0,25 0,016 0.008-0.016 0,25 Compound77K salt 1 0, 125 4 0,25 0,016 0,016 0,5 Compound 78K salt 0,50.064-0.125 2 0, 125 0,008 0,008 0, 125 Compound 79K salt 0,5 0,016 20,064 ≦0.004 ≦0.004 0, 125 Compound 80K salt 1 0,064 4 0,25 ≦0.004≦0.004 0, 125 Compound 81K salt 0,25 0,064 2 0, 125 ≦0.004 ≦0.004 0,064Compound 82K salt 4 0.125-0.25 8 1 0,016 0,016 1 Compound 83K salt 0,125 0,016 0,5 0,032 ≦0.004 ≦0.004 0,064 Compound 84K salt 8 0.125-0.25 82 0, 125 0, 125 2 Compound 85K salt 0,25 0,032 1 0,064 ≦0.004 ≦0.0040.064-0.125 Compound 86K salt 2 0, 125 8 0.5-1 0.016-0.032 0,016 0,5Compound 87K salt 4 0, 125 8 0,25 0,064 0,064 8 Compound 88K salt 1 0,125 4 0,5 0,008 0,008 0,5 Compound 89K salt 4 0, 125 8 0,5 0,032 0,032 2Compound 90K salt 2 0, 125 4 0,5 0,008 0,008 0,25 Ciprofloxacin 4 0,0080,25 1 0,25 0,5 >8 Levofloxacin 4 0,016 1 0,5 0, 125 0,25 8 Doxycycline2 1 >8 0, 125 0, 125 0,25 4

In addition, specific MICs on K. pneumoniae for compound 2, compound 33and compound 35 are shown in Table 14.

TABLE 14 showing MIC on K. pneumoniae for a selection of4-oxoquinolizines. MICs (μg/mL) K. pneumoniae K. pneumoniae BAA-2146Compounds Structure ATCC 33495 (NDM-1, FQ-R) Compound 33 K salt

0, 064 >8 Compound 35 K salt

0, 125 >8 Compound 2 K salt

0, 25 >8

Example 6: Synergy of Antimicrobial Activity of 4-Oxoquinolizines withPolymyxin B Against Acinetobacter and K. pneumonia Strains

Example 6 shows that 4-oxoquinolizine compounds possess synergisticantibacterial activity with polymyxin B. A chequerboard technique wasused to show synergistic interactions between three 4-oxoquinolizinescompounds and polymyxin B (Table 15 and 16, FIGS. 1, 2 and 3). There isa strong indication that compounds 2, 33 and 35 in combination withpolymyxin B are synergistic (or at least partially synergistic) againstfluoroquinolone-resistant strains of A. baumannii. No interaction wasseen between levofloxacin and polymyxin B.

A chequerboard technique was used to identify possible synergistic orantagonistic interactions between the 4-oxoquinolizines compounds,levofloxacin and polymyxin B. Levofloxacin was included as a control.10% Aqueous DMSO stock solutions were prepared for compound 2 (25.6mg/mL), Compound 33 (12.8 mg/mL), and compound 35 (12.8/mL) as well as alevofloxacin stock solution of 1.28 mg/mL and a polymyxin B stocksolution of 0.64 mg/mL. Final dilutions were made in Mueller-Hintonbroth, the specific test medium used for MIC determinations according toCLSI guidelines (CLSI. Methods for Dilution Antimicrobial SusceptibilityTests for Bacteria That Grow Aerobically; Approved Standard-EighthEdition (2009). CLSI Document M07-A8. CLSI, Wayne, Pa. 19087-1898,USA.). The test medium used was a cation-adjusted Mueller-Hinton BrothII (Becton Dickinson UK Ltd., Oxford Science Park, Oxford, UK, OX4 4DQ).

The compounds 2, 33 and 35, as well as levofloxacin were tested incombination with polymyxin B on the following isolates: GN31Acinetobacter baumannii—antibiotic susceptible clinical isolate, GN52Acinetobacter baumannii—levofloxacin resistant clinical isolate, GN56Acinetobacter baumannii—levofloxacin resistant clinical isolate, GN48Klebsiella pneumoniae—NCTC 13443—NDM-1-metallo-β-lactamase and GN10Pseudomonas aeruginosa—ATCC 27853—antibiotic susceptible referenceisolate. All isolates are from the collection maintained at QuotientBioresearch Ltd., Cambridge, UK.

The MIC values for the compounds and polymyxin B and levofloxacin weredetermined by broth microdilution following CLSI guidelines, and theMICs of agents in combination were determined as described by Pillai et.al (Pillai S K, Moellering R C Jr, Eliopoulis G M; AntimicrobialCombinations in Antibiotics in Laboratory Medicine 5^(th) Edition (V.Lorian Ed) (2005) p 365-440). Initial MIC results were used to determinethe microtiter plate patterns for the chequerboards. A suitable doublingdilution concentration range was selected such that the combinationantibiotic range was at least two concentrations above and fourconcentrations below the MIC for each isolate. A fixed range of 0.008 to8 mg/L was tested for polymyxin B except in the case of A. baumanniiGN31 where a range of 0.004 to 4 mg/L was tested when in combinationwith the compounds 2, 33 and 35 as well as levofloxacin.

From the raw data, fractional inhibitory concentration indices (FICI)were determined for the compounds of the invention and levofloxacintogether with polymyxin B for each isolate as follows:

FICI=FIC_(X)+FIC_(Y)

Where,

FIC_(X)=concentration of the combination antibiotic in a particularrow+MIC of the combination drug aloneFIC_(Y)=MIC of polymyxin B in combination+MIC of the polymyxin B alone

Data was interpreted according to the methodology of Pillai: Synergy(FICI<0.5), partial synergy/addition (FICI 0.51 to 0.75), indifference(FICI 0.76 to 2.75), antagonism (FICI>2.75). Additionally isobologramswere created to represent the same data visually. Here, the MIC ofpolymyxin B was plotted against every concentration of the combinationantibiotic up to the MIC. Initial MIC results for all isolates are givenin Table 15 and FICI data are given in Table 16 and Isobolograms areshown in FIGS. 1 to 3.

There is a strong indication that compounds 2, 33 and 35 in combinationwith polymyxin B are synergistic (or at least partially synergistic)against fluoroquinolone-resistant strains of A. baumannii. Nointeraction was seen between levofloxacin and polymyxin B in any of thechequerboard assays performed, all FICI values indicate indifference. Nointeraction was seen in any of the combinations tested for theantibiotic susceptible P. aeruginosa GN10. Surprisingly, also partialsynergy was observed with compound 35 and compound 2 combined withpolymyxin B against the NDM-1-beta-lactamase producing Klebsiellapneumoniae when combined with polymyxin B.

TABLE 15 Pre-chequerboard MIC of compounds for synergy study. MIC mg/LGN31 GN52 GN56 GN48 GN10 Antimicrobial Acinetobacter AcinetobacterAcinetobacter Klebsiella Pseudomonas agent baumannii baumannii baumanniipneumoniae aeruginosa Compound 33 0.03 2 16 16 0.5 Compound 35 0.03 2 816 0.25 Compound 2 0.06 8 16 32 1 Levofloxacin 0.12 8 32 64 1 PolymyxinB 0.5 0.5 0.5 1 1

TABLE 16 FICI data of combinations of Compound 33, 35 and 2 withpolymyxin B Combination Antibiotic in drug Polymyxin B combinationconcentration Combination MIC in Polymyxin B with in combination drugMIC combination MIC alone Isolate polymyxin B (mg/L) alone (mg/L) FICx(mg/L) (mg/L) FICy FICI Synergy GN31 Compound 35 0.004 0.03 0.13 0.120.25 0.48 0.61 Partial synergy Compound 2 0.015 0.06 0.25 0.06 0.25 0.240.49 Synergy Levofloxacin 0.008 0.06 0.13 0.5 0.5 1.00 1.13 IndifferentGN52 Compound 33 1 4 0.25 0.12 0.5 0.24 0.49 Synergy Compound 35 0.5 20.25 0.12 0.5 0.24 0.49 Synergy Compound 2 1 4 0.25 0.12 0.5 0.24 0.49Synergy Levofloxacin 1 8 0.13 0.5 0.5 1.00 1.13 Indifferent GN56Compound 33 2 32 0.06 0.12 0.5 0.24 0.30 Synergy Compound 35 0.5 8 0.060.25 0.5 0.50 0.56 Partial synergy Compound 2 1 16 0.06 0.25 0.5 0.500.56 Partial synergy Levofloxacin 4 32 0.13 0.5 0.5 1.00 1.13Indifferent GN48 Compound 35 1 16 0.06 0.25 0.5 0.50 0.56 Partialsynergy Compound 2 4 32 0.13 0.25 0.5 0.50 0.63 Partial synergyLevofloxacin 8 64 0.13 0.5 0.5 1.00 1.13 Indifferent GN31-Acinetobacterbaumannii —antibiotic susceptible isolate GN52-Acinetobacter baumannii—levofloxacin resistant clinical isolate GN56-Acinetobacter baumannii—levofloxacin resistant clinical isolate GN48-Klebsiella pneumoniae—NCTC 13443-NDM-1-β-lactamase producing isolate

Example 7 Cytotoxicity

Cytotoxicity was determined for the instant 4-oxoquinolizine compoundswhich show low cytotoxicity. (see Table 17 following). Thus, while theinstant 4-oxoquinolizine compounds possess potent activity against bothGram-positive and Gram-negative resistant strains, including nosocomialstrains as well as CDC pathogens, they have good drug profiles regardingsafety and efficacy.

TABLE 17 showing cytotoxity values for the instant 4-oxoquinolizinesCyto- toxicity Com- IC₅₀ pounds Structures (μM) 1

500 2

250 3

50 4

60 7

150 8

90

Example 8: Preparation of 2-Pyridones Compounds Analytical Methods

NMR spectra were recorded on a Bruker Avance-400 NMR or BrukerAvance-300 NMR or with samples in solution in deuterated chloroform(CDCl₃), deuterated MeOH (CD₃OD) or deuterated dimethyl sulfoxide(DMSO-d6). Chemical shifts and coupling constants are respectivelyexpressed in part per million (ppm) and in Herz (Hz). Mass spectrometry(MS) analyses were performed on an Agilent MSD G1946D or a Waters TQDwith electrospray ionization (ESI). High resolution mass spectrometry(High-Res MS) analyses were recorded on a Shimadzu IT-TOF apparatus.HPLC analyses were performed on columns Waters XBridge (C18, 30×2.1 mm,3.5 micron) at a column temperature of 35° C. with a flow rate of 1mL/min of a mixture of eluent A (0.1% Formic acid in ACN) and eluent B(0.1% Formic acid in water); 3 methods of elution were used, method 1,method 2 and method 3 as described below.

HPLC Method 1

Lin. Gradient: t=0 min 2% A, t=1.6 min 98% A, t=3 min 98% A

Detection: DAD (220-320 nm)

Detection: MSD (ESI pos/neg) mass range: 100-800Detection: ELSD (PL-ELS 2100) gas flow 1.1 mL/min; gas temp: 50° C.

HPLC Method 2

Lin. Gradient: t=0 min 2% A, t=3.5 min 98% A, t=6 min 98% A

Detection: DAD (220-320 nm)

Detection: MSD (ESI pos/neg) mass range: 100-800Detection: ELSD (PL-ELS 2100) gas flow 1.1 mL/min; gas temp: 50° C.

HPLC Method 3

Lin. Gradient: t=0 min 2% A, t=10 min 98% A, t=14 min 98% A

Detection: DAD (220-320 nm)

Detection: MSD (ESI pos/neg) mass range: 100-800Detection: ELSD (PL-ELS 2100) gas flow 1.1 mL/min; gas temp: 50° C.

The following acronyms and abbreviations are used:

ACN Acetonitrile BOC t-butoxycarbonyl DCM Dichloromethane DMFN,N-dimethylformamide DMSO Dimethylsulfoxide ESI-MS electrosprayionization mass spectrometry HPLC High-performance liquid chromatographyLCMS Liquid chromatography-mass spectrometry MeOH methanol nBuLi n-butyllithium NMR Nuclear magnetic resonance TFA Trifluoroacetic acid THFTetrahydrofuran

The 2-pyridones compounds were obtained in 2 steps from a suitablescaffold. Most of the compounds were prepared via a Palladium couplingbetween the ester-protected scaffold and a boronate reagent followed byhydrolysis of the ester moiety. Depending of the boronate reagentsadditional deprotection steps could be required. Some compounds werealso made by substituting the scaffolds with an amine instead of apalladium coupling with a boronate hence forming an N—C bond instead ofa C—C bond.

FIG. 4 shows the structure of 5 scaffolds.

Preparation of Scaffolds:

Scaffolds ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate(Scaffold A), methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate(Scaffold B) and methyl8-chloro-1-cyclopropyl-9-methoxy-4-oxo-quinolizine-3-carboxylate(Scaffold E) were prepared synthetically. Scaffolds ethyl8-chloro-1-cyclopropyl-7,9-dimethyl-4-oxo-quinolizine-3-carboxylate(Scaffold D) was obtained as a side product of the synthesis of scaffoldB. Scaffold methyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylate(Scaffold C) was purchased from Beijing Louston Fine Chemical Co. Ltd.,China.

Scaffolds A, B and E were prepared in 6-7 steps from commercial2-bromo-3-methyl-4-chloro-pyridine.

Preparation of scaffold A: ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylatePreparation of 2-bromo-3-methyl-4-chloro-pyridine

To a solution of 2,2,6,6-tetramethyl-pyridine (21.1 mL, 125 mmol) infreshly distilled THF (120 mL) at −78° C. was added nBuLi (50 mL, 125mmol) in 30 min. The resulting mixture was stirred at −78° C. for 30 minand was added through a cannula over 30 min to a solution of3-bromo-4-chloro-pyridine (20.0 g, 104 mmol) in freshly distilled THF(60 mL) that had been cooled to −78° C. prior to the addition. Thereaction mixture was stirred at −78° C. for 30 min before iodomethane(7.78 mL, 125 mmol) was added over a period of 10 min. The reaction wasstirred at −78° C. for 30 min and was allowed to warm up to roomtemperature prior to be quenched with aqueous NH₄Cl (65 mL). The aqueousphase was extracted with ethyl acetate (2×150 mL). The organic phaseswere separated, dried, and concentrated. The residue was purified byflash silica column chromatography (hexane:ethyl acetate, 5:1) to affordthe title compound as a yellow solid (10.6 g, 49%).

¹H NMR (CDCl₃, 300 MHz) 5 ppm: 8.10 (d, J=5.1 Hz, 1H), 7.27 (d, J=5.1Hz, 1H), 2.51 (s, 3H).

Preparation of (3-methyl-4-chloro-pyridin-2-yl)-cyclopropyl-methanol

A solution of 2-bromo-3-methyl-4-chloro-pyridine (10.6 g, 57.1 mmol) infreshly distilled THF (120 mL) was cooled down to 0° C. and treated withisopropyl magnesium chloride (45.7 mL, 2.0 M in THF, 91.5 mmol). Theresulting mixture was stirred at room temperature for 3 h then cooled to−5° C. Cyclopropane carboxaldehyde (6.83 mL, 91.5 mmol) was added. Thereaction mixture was stirred at room temperature for 1 h and quenched byadding water (100 mL), and extracted with ethyl acetate (2×150 mL). Theorganic phase was separated, dried, and concentrated. The residue waspurified by flash silica column chromatography (hexane:ethyl acetate,3:1) to afford the title compound as a yellow oil (7.01 g, 62%).

ESI-MS m/z: 198 (M+H)⁺; ¹H NMR (CDCl₃, 300 MHz) δ ppm: 8.28 (d, J=5.4Hz, 1H), 7.26 (d, J=5.4 Hz, 1H), 4.79 (d, J=5.4 Hz, 1H), 4.55 (br s,1H), 2.39 (s, 3H), 1.10-1.28 (m, 1H), 0.41-0.58 (m, 4H).

Preparation of (3-methyl-4-chloro-pyridin-2-yl)-cyclopropyl-methanone

A solution of (3-methyl-4-chloro-pyridin-2-yl)-cyclopropyl-methanol(7.01 g, 35.5 mmol) in DCM (80 mL) was treated with MnO₂ (30.8 g, 355mmol) at room temperature overnight. The reaction mixture was filteredthrough a pad of Celite and the filtrate was concentrated to dryness,affording the title compound (6.82 g, 98%).

ESI-MS m/z: 196 (M+H)⁺; ¹H NMR (CDCl₃, 300 MHz) δ ppm: 8.41 (d, J=5.4Hz, 1H), 7.42 (d, J=5.4 Hz, 1H), 2.96-3.04 (m, 1H), 2.57 (s, 3H),1.20-1.28 (m, 2H), 1.09-1.14 (m, 2H).

Preparation of2-(1-cyclopropyl-2-methoxy-vinyl)-3-methyl-4-chloro-pyridine

A solution of methoxymethyl triphenylphosphonium chloride (17.9 g, 52.3mmol) in dry THF (80 mL) was treated with NaH (2.79 g, 69.8 mmol) at 0°C. for 3 h. To this mixture was added a solution of(3-methyl-4-chloro-pyridin-2-yl)-cyclopropyl-methanone (6.82 g, 34.9mmol) in dry THF (20 mL). The reaction mixture was heated at 40° C.overnight. The reaction mixture was cooled down to room temperature andfiltered. The filtrate was concentrated to dryness. The residue waspurified by flash silica column chromatography (hexane:ethyl acetate,3:1) to afford the title compound (6.41 g, 82%).

ESI-MS m/z: 224 (M+H)⁺; ¹H NMR (CDCl₃, 300 MHz) δ ppm: 8.31 (d, J=5.4Hz, 0.5H), 8.23 (d, J=5.4 Hz, 0.5H), 7.17 (d, J=5.4 Hz, 1H), 6.13-6.16(m, 1H), 3.70 (s, 1.5H), 3.56 (s, 1.5H), 2.38 (s, 1.5H), 2.31 (s, 1.5H),1.91-1.94 (m, 0.5H), 1.63-1.65 (m, 0.5H), 0.66-0.72 (m, 1H), 0.56-0.62(m, 1H), 0.35-0.38 (m, 1H), 0.26-0.33 (m, 1H).

Preparation of2-(3-methyl-4-chloro-pyridin-2-yl)-2-cyclopropyl-acetaldehyde

A solution of2-(1-cyclopropyl-2-methoxy-vinyl)-3-methyl-4-chloro-pyridine (6.41 g,28.7 mmol) in acetic acid (50 mL) was treated with sulfuric acid (6.52mL, 143 mmol) at room temperature overnight. The reaction mixture wasneutralized with 2N NaOH to pH 8-9 extracted with ethyl acetate (2×100mL). The organic phases were combined, dried, and concentrated. Theresidue was purified by flash silica column chromatography (hexane/ethylacetate=2:1) to afford the title compound as a yellow solid (4.83 g,80%).

ESI-MS m/z: 210 (M+H)⁺; ¹H NMR (CDCl₃, 300 MHz) δ ppm: 9.89 (d, J=2.4Hz, 1H), 8.35 (d, J=5.1 Hz, 1H), 7.26 (d, J=5.4 Hz, 1H), 3.26-3.28 (m,1H), 2.35 (s, 3H), 1.53-1.59 (m, 1H), 0.55-0.79 (m, 2H), 0.25-0.39 (m,2H).

Preparation of2-[2-(3-methyl-4-chloro-pyridin-2-yl)-2-cyclopropyl-ethylidene]-malonicacid diethyl ester

A mixture of (3-methyl-4-chloro-pyridin-2-yl)-cyclopropyl-acetaldehyde(4.83 g, 23.0 mmol), diethyl malonate (7.02 g, 43.8 mmol), piperidine(3.62 mL, 36.6 mmol), and acetic acid (4.19 mL, 73.2 mmol) in ethanol(100 mL) was heated to reflux overnight. The reaction mixture wasconcentrated to dryness. The residue was purified by flash silica columnchromatography (hexane:ethyl acetate, 4:1) to afford the title compoundas a yellow oil (5.76 g, 71%).

ESI-MS m/z: 352 (M+H)⁺.

Preparation of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A solution of2-[2-(3-methyl-4-chloro-pyridin-2-yl)-2-cyclopropyl-ethylidene]-malonicacid diethyl ester (5.76 g, 16.4 mmol) in Dowtherm A (80 mL) was heatedin a preheated oil bath at 230° C. for 15 min. The reaction mixture wascooled to room temperature and purified by flash silica columnchromatography (hexane:ethyl acetate, 2:1 to 1:2) to afford the titlecompound as a yellow solid (3.76 g, 75%).

ESI-MS m/z: 306 (M+H)⁺; ¹H NMR (CDCl₃, 300 MHz) 5 ppm: 9.34 (d, J=7.8Hz, 1H), 8.40 (s, 1H), 7.12 (d, J=7.8 Hz, 1H), 4.41 (q, 2H), 3.00 (s,3H), 2.28-2.33 (m, 1H), 1.42 (t, 3H), 1.03-1.08 (m, 2H), 0.71-0.76 (m,2H).

Preparation of scaffold B: methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylatePreparation of 2-bromo-4-chloro-3-methyl-pyridine

In a 250 mL 3-neck flask, 2,2,6,6-tetramethyl-pyridine (10.5 mL, 61.7mmol) was added to dry THF (100 mL) under inert atmosphere and theresulting yellow solution was cooled down to −78 OC. N-Butyl lithium 2.5M solution in hexane (25 mL, 62.5 mmol) was added over 0.5 h and thereaction mixture was stirred at −78 OC for 0.5 h. The solution hadturned from yellow to orange.

In a 500 mL 3-neck flask, 3-bromo-4-chloro-pyridine (9.6 g, 49.9 mmol)was added to dry THF (50 mL) under inert atmosphere. This reactionmixture was cooled down to −78 OC. The orange solution previouslyobtained was added via cannula over 0.75 h and stirred at −78° C. for0.5 h. The solution/suspension turned black. Iodomethane (3.9 mL, 62.6mmol) was added over 0.5 h and the reaction mixture was stirred at −78OC for 0.5 h. The reaction mixture was left to warm to room temperatureand was quenched by addition of an aqueous saturated ammonium chloridesolution (65 mL). The reaction mixture was diluted with water (100 mL),ethanol (100 mL) and ethyl acetate (100 mL). The mixture was extractedwith ethyl acetate (2×100 mL). The organic layers were combined andwashed with 50% brine (1:1 water/brine, 100 mL total) and brine (100mL). Combined water layers were extracted with ethyl acetate (100 mL).The combined organic layers were dried over sodium sulfate, filtered andevaporated to dryness to yield a brown oil.

The crude oil was dissolved again in ethyl acetate (50 mL), and ammoniumsalts precipitated and were filtered off. The filtrate was evaporated todryness to yield a brown oily solid. A third of the obtained crudeproduct was distillated with a Kügelrohr apparatus at 100° C. under a0.1 mbar reduced pressure. The distillate crystallized out on coolingyielding the intended compound (1.68 g, 12.4%). The remaining crudeproduct was purified by flash chromatography using a gradient of 10-75%ethyl acetate in heptane affording a second batch of the compound (3.53g, 24.3). The combined batches yielded the intended product (5.21 g,36.7%).

¹H NMR (400 MHz, CDCl₃) δ ppm 8.11 (d, J=5.2 Hz, 1H), 7.27 (d, J=6 Hz,1H), 2.52 (s, 3H).

Preparation of (4-chloro-3-methyl-2-pyridyl)-cyclopropyl-methanol

A solution of 2-bromo-4-chloro-3-methylpyridine (7 g, 28.1 mmol) in dryTHF (150 mL) was cooled down to 0° C. An isopropyl magnesiumchloride—Lithium chloride complex (26 mL, 33.8 mmol) was added carefullyso temperature would not rise above 5° C. during the addition. Themixture was allowed to warm up to room temperature and was stirred for 1h. The reaction mixture was cooled down to 0° C. and cyclopropanecarboxaldehyde (2.5 mL, 33.5 mmol) was added carefully so temperaturewould not rise above 5° C. during the addition. The mixture was allowedto warm up to room temperature and was stirred for 1 h. The reactionmixture was cooled to 0° C. and water (150 mL) was added carefully sotemperature would not rise above 10° C. during the addition. The mixturewas allowed to warm up to room temperature and was stirred for 1 h. Themixture was extracted with ethyl acetate (3×100 mL). The combinedorganic phases were washed with brine (100 mL), dried over sodiumsulfate and evaporated. The residue was purified by flash chromatographyover silica gel (0-30% ethyl acetate in heptane) yielding a yellow oil(3.47 g, 16.8%).

LC-MS: t=1.33 min (method 1); 198 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm8.28 (d, J=5.2 Hz, 1H), 7.25 (d, J=5.6 Hz, 1H), 4.80 (dd, J=7.8 and 5.3Hz, 1H), 4.51 (d, J=5.6 Hz, 1H), 2.38 (s, 3H), 1.05-1.15 (m, 1H),0.52-0.58 (m, 1H), 0.44-0.50 (m, 1H), 0.35-0.42 (m, 1H).

Preparation of (4-chloro-3-methyl-2-pyridyl)-cyclopropyl-methanone

Manganese dioxide (1126 mg, 12.95 mmol) was added to a solution of(4-chloro-3-methylpyridin-2-yl)-cyclopropyl-methanol (250 mg, 1.214mmol) in DCM (10 mL). The reaction mixture was stirred at roomtemperature for 45 h. The mixture was filtered over a porosity 4 filter.The filtrate was concentrated to dryness to yield white crystals (245mg, 98%).

LC-MS: t=1.92 min (method 1); 196 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm8.41 (d, J=5 Hz, 1H), 7.44 (d, J=5.3 Hz, 1H), 5.30 (s, 1H), 2.98-3.05(1H, m), 2.52 (s, 3H), 1.24-1.29 (m, 2H), 1.08-1.14 (m, 2H).

Preparation of4-chloro-2-[1-cyclopropyl-2-methoxy-vinyl]-3-methyl-pyridine

A yellow suspension of (4-chloro-3-methylpyridin-2-yl) (cyclopropyl)methanone (2.99 g, 15.28 mmol), (Methoxymethyl) triphenylphosphoniumchloride (7.84 g, 22.87 mmol) and Potassium tert-butoxide (3.41 g, 30.4mmol) in toluene (50 mL) was heated to 60° C. and stirred for 3.5 h. Thereaction mixture was cooled down to room temperature and an aqueoussolution of 4M Hydrochloric acid (50 mL) was added. The reaction mixturewas washed with toluene (3×50 mL). The aqueous layer was diluted in ice,and solid sodium bicarbonate was added until pH reaches 7-8. The mixturewas extracted with ethyl acetate (3×100 mL). The combined organic layerswere washed with brine, dried over sodium sulfate, filtrated andevaporated to dryness. The oily residue was purified by flash columnchromatography over silica gel using a gradient of ethyl acetate inheptane to yield a 1 to 1 mixture of the isomeric vinyl ethers as yellowoil (2.93 g, 82%).

LC-MS: t=1.68 min and 1.73 min (E and Z isomers) (method 1); 224 (M+H)⁺.

Preparation of2-(4-chloro-3-methyl-2-pyridyl)-2-cyclopropyl-acetaldehyde

To a solution of4-chloro-2-(1-cyclopropyl-2-methoxyvinyl)-3-methylpyridine (1.7 g, 7.60mmol) in THF (15 mL) a 2M aqueous solution of sulfuric acid (15.20 mL,30.4 mmol) was added and the reaction mixture was stirred at 50° C. fora total of 2.5 h. The resulting mixture was poured into water andneutralized with a saturated sodium bicarbonate solution, andthereafter, extracted with chloroform (3×20 mL). The resulting organiclayers were washed with saturated salt water (20 mL), dried over sodiumsulfate and concentrated under reduced pressure. The resulting yellowsolid was purified by silica gel column chromatography (hexane:ethylacetate) (4:1) to obtain the aldehyde (1.08 g, 67.8%).

LC-MS: t=1.55 min (method 1); 210 (M+H)⁺; 228 (M+H₂O+H)⁺.

Preparation of methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate

Piperidine (7 mL, 70.9 mmol) and acetic acid (5.5 mL, 95 mmol) wereadded to a solution of2-(4-chloro-3-methylpyridin-2-yl)-2-cyclopropylacetaldehyde (4.98 g,23.75 mmol) in absolute ethanol (200 mL). Dimethyl malonate (16.44 mL,144 mmol) was added and the reaction mixture was stirred at 100° C. for5 h (the reaction mixture turned into a red solution). The solvent wasevaporated under reduced pressure. The resulting mixture was dilutedwith ether (100 mL) and washed with water (100 mL) and brine (50 mL).The organic layer was separated and dried over sodium sulfate. Themixture was evaporated to dryness. Dowtherm A (110 mL) was added. Thisreaction mixture was heated to 240° C. under microwave irradiation andstirred at this temperature for 0.5 h during which the reaction mixtureturned into a black solution. The residue was purified by reversed phaseflash chromatography using a 5%-100% ACN gradient in water with 1% TFAyielding the cyclized methyl ester (4 g, 51.4%).

LC-MS: t=1.88 min (method 1); 292 (M+H)⁺.

Preparation of scaffold D: methyl8-chloro-1-cyclopropyl-7,9-dimethyl-4-oxo-quinolizine-3-carboxylate

During the first scale up preparation of methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate a sideproduct was observed in the first step. Other scale-up preparations ofthe scaffold were optimized to avoid the side product. The side productwas carried over till the last step. Reversed phase purification (40-80%ACN in 0.1% formic acid in water over C₁₋₈-silica) afforded methyl8-chloro-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylateas a yellow solid (0.4 g).

ESI-MS m/z: 306 (M+H)⁺; 1H NMR (400 MHz DMSO-d6) δ ppm 9.33 (s, 1H),8.36 (s, 1H), 3.93 (s, 3H), 3.04 (s, 3H), 2.48 (s, 3H), 2.27-2.37 (m,1H), 1.01-1.09 (m, 2H), 0.70-0.77 (m, 2H); 13C NMR (100 MHz CDCl₃) δ ppm166.7, 155.2, 147.1, 145.1, 144.0, 130.9, 127.5, 125.7, 115.7, 104.6,52.1, 20.0, 18.5, 17.0, 9.76 (2 C).

Preparation of scaffold E: methyl8-chloro-1-cyclopropyl-9-methoxy-4-oxo-quinolizine-3-carboxylatePreparation of 2-bromo-4-chloropyridin-3-ol

A solution of 2.5 M n-BuLi in hexanes (180 mL, 450 mmol) was addeddropwise to a solution of 2,2,6,6-Tetramethylpiperidine (75 mL, 441mmol) in THF (800 mL) at −70° C. under inert atmosphere. The reactionmixture was agitated for 2 h and transferred to a solution of3-bromo-4-chloropyridine (80.5 g, 418 mmol) in THF (500 mL) at −70° C.under inert atmosphere and stirred for 2 h. Trimethyl borate (100 mL,881 mmol) was added dropwise and reacted for 2 h. 33% Peracetic acid inacetic acid (150 mL, 780 mmol) was added dropwise, the mixture waswarmed to room temperature and stirred for 14 h. The mixture was cooledto 0° C. and sodium metabisulfite (200 g, 1.05 mol) in water (400 mL)was added over 2 h. Water (300 mL) and ethyl acetate (200 mL) wereadded. The layers were separated and the aqueous layer washed with ethylacetate (3×1000 mL). The organic phase was dried over sodium sulfate,filtered and evaporated to dryness to obtain crude product which waspurified over silica gel, using 0:1 to 3:2 ethyl acetate in heptane) toobtain 2-bromo-4-chloropyridin-3-ol as a white solid (46.3 g, 50%).

ESI-MS m/z: 210 (M+H)⁺.

Preparation of 2-bromo-4-chloro-3-methoxy-pyridine

2-bromo-4-chloropyridin-3-ol (8.15 g, 39.1 mmol), potassium carbonate(10.5 g, 76 mmol) and iodomethane (3.65 ml, 58.6 mmol) were added toacetone (300 ml) and stirred for 18 h at room temperature under inertconditions. The reaction mixture was evaporated to dryness, dissolved inethyl acetate (100 mL), filtered over silica gel and the filter washedwith ethyl acetate (3×100 mL). The filtrate was evaporated to dryness toobtain crude product which was purified over silica gel using 0:1 to 1:0ethyl acetate in heptane to obtain 2-bromo-4-chloro-3-methoxypyridine asa white crystalline solid (6.8 g, 78%).

ESI-MS m/z: 224 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 8.06 (d, J=3.6 Hz,1H), 7.31 (d, J=5.0 Hz, 1H), 3.95 (s, 1H).

Preparation of (4-chloro-3-methoxy-2-pyridyl)-cyclopropyl-methanol

2-bromo-4-chloro-3-methoxy-pyridine (2.97 g, 13.4 mmol) was added to THF(dry) (100 ml) and cooled to 0° C. Isopropylmagnesium chloride—lithiumchloride complex (13.5 ml, 17.6 mmol) was added and the reaction mixturestirred for 0.5 h at room temperature. The mixture was cooled to 0° C.,cyclopropanecarboxaldehyde (1.297 ml, 17.4 mmol) was added and thereaction mixture stirred for 1 h at room temperature. The mixture wascooled to 0° C., water (72.3 ml, 4 mol) was added and the reactionmixture stirred overnight at room temperature. The reaction mixture wasextracted with ethyl acetate (3×75 mL), the organic phase dried oversodium sulfate, filtered and evaporated to dryness to obtain crudeproduct which was purified over silica gel using 0:1 to 1:0 ethylacetate in heptane to obtain(4-chloro-3-methoxy-2-pyridyl)-cyclopropyl-methanol as a yellow oil (2.4g, 70%) ESI-MS m/z: 214 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 8.23 (d,J=5.0 Hz, 1H), 7.29 (d, J=5.0 Hz, 1H), 4.63 (t, J=7.8 Hz, 1H), 4.00 (d,J=8.1 Hz, 1H), 3.93 (s, 3H), 1.15-1.22 (m, 1H), 0.57-0.63 (m, 1H),0.42-0.50 (m, 3H).

Preparation of (4-chloro-3-methoxy-2-pyridyl)-cyclopropyl-methanone

To a solution of (4-chloro-3-methoxy-2-pyridyl)-cyclopropyl-methanol(700 mg, 3.28 mmol) in DCM (20 ml), manganese (IV) oxide (4.6 g, 58mmol) was added and stirred for 14 h at room temperature. The reactionmixture was filtered over Celite and the filtrate evaporated to drynessto obtain (4-chloro-3-methoxy-2-pyridyl)-cyclopropyl-methanone as ayellow oil (668 mg, 96%).

ESI-MS m/z: 212 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 8.35 (d, J=5.0 Hz,1H), 7.49 (d, J=5.0 Hz, 1H), 3.95 (s, 3H), 2.96-2.89 (m, 1H), 1.28-1.33(m, 2H), 1.08-1.16 (m, 2H).

Preparation of4-chloro-2-(1-cyclopropyl-2-methoxyvinyl)-3-methoxypyridine

(Methoxymethyl)triphenylphosphonium chloride (1.6 g, 4.7 mmol) wasdissolved in THF (20 ml) and cooled to −30° C. 2.5 Molar n-BuLi inhexanes (1.89 ml, 4.7 mmol) was and the mixture stirred for 1 h.(4-chloro-3-methoxypyridin-2-yl)-(cyclopropyl)methanone (500 mg, 2.4mmol) dissolved in THF (5 ml) was added slowly and stirred for 4 h. Thereaction mixture was quenched with saturated ammonium chloride (20 mL)and stirred overnight. 25 mL ethyl acetate was added and the layerspartitioned. The organic phase was dried over sodium sulfate, filteredand evaporated to dryness to obtain crude product which was purifiedover silica gel using 0:1 to 3:7 ethyl acetate in heptane to obtain4-chloro-2-(1-cyclopropyl-2-methoxyvinyl)-3-methoxy-pyridine as a clearoil (361 mg, 63%).

ESI-MS m/z: 240 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 8.16 (d, J=5.3 Hz,1H), 7.17 (d, J=5.0 Hz, 1H), 6.57 (s, 1H), 3.82 (s, 3H), 3.76 (s, 3H),1.83-1.92 (m, 1H), 0.70-0.77 (m, 2H), 0.44-0.51 (m, 2H).

Preparation of2-(4-chloro-3-methoxypyridin-2-yl)-2-cyclopropyl-acetaldehyde

4-chloro-2-(1-cyclopropyl-2-methoxyvinyl)-3-methoxy-pyridine (259 mg,1.1 mmol) was dissolved in THF (10 ml) and cooled to 0° C. 4M sulfuricacid (2.7 ml, 10 mmol) was added and the reaction was performed atreduced pressure while heating at 50° C. for 2.5 h. The mixture wasdiluted in ice water (10 mL) and sodium bicarbonate added untilneutralized. The mixture was washed with DCM (3×10 mL) and the organicphase dried, filtered and evaporated to dryness to obtain2-(4-chloro-3-methoxy-pyridin-2-yl)-2-cyclopropyl-acetaldehyde as ayellow oil (233 mg) to be used without further purification.

ESI-MS m/z: 226 (M+H)⁺.

Preparation of methyl8-chloro-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate

2-(4-chloro-3-methoxy-pyridin-2-yl)-2-cyclopropyl-acetaldehyde (4.13 g,18.3 mmol) was dissolved (60 ml). Acetic acid (4.2 ml, 73 mmol),piperidine (3.6 ml, 37 mmol) and dimethyl malonate (12.5 ml, 110 mmol)were added and stirred at 100° C. for 5 h. The solvent was distilled offin vacuum, the resulting mixture diluted with diethyl ether (200 mL),washed with water (100 mL) and brine (100 mL). The organic phase wasseparated, dried over sodium sulfate, filtered and evaporated to drynessto obtain a red oil which was not purified. The mixture was dissolved inDowtherm A (100 ml) and heated at 240° C. for 1 h. The reaction mixturewas purified over silica gel, rinsing off the Dowtherm A with heptane.The product was purified using 0:1 to 1:0 ethyl acetate in heptane toobtain: methyl8-chloro-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate asan orange-yellow crystalline solid (2.83 g, 47%).

ESI-MS m/z: 308 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 9.27 (d, J=7.8 Hz,1H), 8.28 (s, 1H), 7.13 (d, J=7.8 Hz, 1H), 3.98 (s, 3H), 3.94 (s, 3H),2.47-2.56 (m, 1H), 0.97-1.04 (m, 2H), 0.73-0.78 (m, 2H).

Preparation of Compounds 1-17 from Scaffold B

The compounds were first prepared according to the following methods.The preparation of compounds 1, 2, 3, 4, 5, 6, and 17 is describedbelow. The same methods were also used to prepare compounds 7-16.Chemical names and structures of the compounds are given in Table 1.

Preparation of Compound 2 Preparation of2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

A 0.4 M solution of 4-Bromo-2,5-difluoroaniline (400 mg, 1.923 mmol) indimethyl sulfoxide (4.8 mL) was added to sodium acetate (473 mg, 5.77mmol) and bis-pinacolato diborane (537 mg, 2.115 mmol) in a flask underargon. The mixture was degassed with argon prior to the addition ofbis-(triphenylphosphine) palladium(II) chloride (13.50 mg, 0.019 mmol).The reaction mixture was heated at 80° C. until complete consumption ofstarting material (16 h). After cooling the reaction mixture to roomtemperature ethyl acetate was added and the reaction mixture waspartitioned between ethyl acetate and a saturated aqueous sodiumbicarbonate solution. The organic layer was washed with brine (4 times)to remove the dimethyl sulfoxide. The material was dried with sodiumsulfate and concentrated in vacuum. The crude product was purified byflash silica column chromatography (heptane/5% ethyl acetate) to give awhite solid (220 mg, 44.9%).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.26-7.30 (m, 1H), 6.38-6.43 (m, 1H), 4.02(s, 2H), 1.33 (s, 12H).

Preparation of methyl8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropy-9-methyl-4-oxo-quinolizine-3-carboxylate

The following Suzuki coupling method was used towards the preparation ofcompound 2 and is referred as the general Suzuki coupling method for thepreparation of the other compounds.

General Suzuki Coupling Method:

Ethanol (96%) (129 μl), 2M aqueous sodium carbonate (175 μl, 0.350mmol),2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(38.6 mg, 0.152 mmol) were added to a solution of methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (34mg, 0.117 mmol) in toluene (250 μl). The mixture was degassed withargon. 1,1′-Bis-diphenylphosphine ferrocene palladium(II) dichloride(8.52 mg, 0.012 mmol) was added and the mixture was heated at 90° C.under an argon atmosphere for 4 h. The reaction mixture was cooled. Themixture was diluted with DCM (3 mL) and water was added (3 mL). Thelayers were separated using a phase separator and the aqueous layer wasextracted with DCM (3×2 mL). The combined organic layers wereconcentrated in vacuum. Purification by with flash silica columnchromatography using a gradient (heptane/ethyl acetate) (2:1 to 1:1)afforded a yellow solid (46 mg, 100%).

LC-MS: t=2.00 min (method 1); 385 (M+H)⁺; 383 (M−H)⁻.

Preparation of compound 28-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A solution of methyl8-(4-amino-2,5-difluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(36 mg, 0.094 mmol) and sodium hydroxide 1N aqueous solution (0.5 mL,0.5 mmol) in MeOH (2 mL) was stirred at 50° C. for 2 h. The reactionmixture was cooled down and the MeOH was removed in vacuum and theresidue was taken-up in 5 mL water and then neutralized with 1NHydrochloric acid (˜0.5 mL). A precipitation was formed and the mixturewas stirred at room temperature overnight. The yellow solid wascollected by filtration and dried in a desiccator. The precipitate wastriturated with toluene/iso-propanol (1:1) (4 mL) and filtrationafforded the product compound 2 (9 mg, 25.9%).

LC-MS: t=2.07 min (method 1); 371 (M+H)⁺; 369 (M−H)⁻; 1H NMR (400 MHz,CDCl₃) b ppm 9.45 (d, J=6 Hz, 1H), 8.43 (s, 1H), 7.05 (d, J=7.3 Hz, 1H),7.05 (d, J=7.3 Hz. 1H), 6.97 (dd, J=11 and 6.3 Hz, 1H), 6.62 (dd, J=10.6and 7.3 Hz, 1H), 5.30 (s, 2H), 2.30-2.40 (m, 1H), 2.05 (s, 3H),1.00-1.10 (m, 2H), 0.75-0.82 (m, 2H).

Preparation of Compound 1 Preparation of compound 18-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

The general Suzuki coupling method described above was used to couplemethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate with3-fluoro-4-butyloxycarbonyl-aminophenyl boronic acid. Purification byflash silica column chromatography yielded methyl8-[4-(tert-butoxycarbonylamino)-3-fluoro-phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylateas a yellow solid. A solution of the obtained solid and TFA (0.2 mL) inDCM (1 mL) was stirred at room temperature for 2 h.

The product was lyophilized and dissolved in a mixture of THF (0.5 mL).An aqueous 4N sodium hydroxide solution (0.33 mL) was added before beingirradiated twice at 120° C. in a microwave oven for 10 min. The productwas purified by preparative HPLC yielding compound 18-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid (19.2 mg, 16%).

LC-MS: t=8.30 min (method 3); 353 (M+H)⁺; 351 (M−H)⁻.

Preparation of Compound 3 Preparation of3,5-dichloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline

A solution of 4-bromo-2,6-dichloroaniline (1 g, 4.15 mmol) in dimethylsulfoxide (10 mL) (0.4 M) was added to sodium acetate (1.022 g, 12.45mmol) and bis-pinacolato diborane (1.159 g, 4.57 mmol) in a flask underargon. The mixture was degassed with argon. 1,1′-Bis-diphenylphosphineferrocene palladium(II) dichloride (0.328 g, 0.415 mmol) was added andthe reaction mixture was heated at 80° C. for 16 h. After cooling toroom temperature ethyl acetate (50 mL) was added and the reactionmixture was filtered and partitioned between ethyl acetate and asaturated aqueous sodium bicarbonate solution (50 mL). The organic layerwas washed with brine (4×50 mL), dried sodium sulfate and concentratedin vacuum. The crude product was purified by flash silica columnchromatography (heptane/5% ethyl acetate) to give a white solid (470 mg,39.3%).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.60 (s, 2H), 4.67 (s, 2H), 1.32 (s, 12H).

Preparation of methyl8-(4-amino-3,5-dichloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate

The general Suzuki coupling method described above was used to couplemethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate with3,5-dichloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline.Purification by flash silica column chromatography yielded methyl8-(4-amino-3,5-dichloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylateas a yellow solid (41 mg, 78%).

LC-MS: t=2.14 min (method 1); 417 (M+H)⁺ and isotopic 419 (M+H)⁺.

Preparation of compound 38-(4-amino-3,5-dichloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A solution of methyl8-(4-amino-3,5-dichlorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(29.2 mg, 0.052 mmol) and sodium hydroxide 1N aqueous solution (0.5 mL,0.5 mmol) in MeOH (2 mL) was stirred at 50° C. for 2 h. The mixture wascooled and the MeOH was evaporated. The residue was taken-up in 5 mLwater and neutralized with 1N hydrochloric acid ˜0.5 mL. A yellowprecipitation was formed and the precipitate was stirred at roomtemperature for 4 h. The yellow solid was collected by filtration anddried in a desiccator over potassium hydroxide. The crude product waspurified with preparative HPLC and freeze-dried yielding compound 3 as ayellow solid (13 mg, 61.4%).

LC-MS: t=3.73 min (method 2); 403 (M+H)⁺ and isotopic 405 (M+H)⁺; 401(M−H)⁻ and isotopic 403 (M−H)⁻.

Preparation of Compound 4 Preparation of tert-butylN-[(4-bromo-2-fluoro-phenyl)methyl]-carbamate

Di-tert-butyl (4.01 g, 18.38 mmol) and triethylamine (2.56 mL, 18.38mmol) were added to a solution of 4-bromo-2-fluorophenyl)-1-methylamine(2.5 g, 12.25 mmol) in DCM (50 mL) at 0° C. The mixture was allowed towarm up to room temperature and stirred for 2 h. Water was added (50 mL)and the layers were separated. The organic layer was washed with brine(3×50 mL), dried with sodium sulfate and concentrated in vacuum. Thecrude product was purified by flash silica column chromatography(heptane:ethyl) acetate (10:0 to 8:2) to yield a colorless oil (2.65 g,71.1%).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.20-7.30 (m, 3H), 4.90 (s, 1H), 4.30 (d,J=6 Hz, 2H), 1.44 (s, 9H).

Preparation of tert-butylN-[[2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl]carbamate

A mixture of tert-butyl N-[(4-bromo-2-fluoro-phenyl)methyl]carbamate(1.3 g, 4.27 mmol), bis-pinacolato diborane (1.628 g, 6.41 mmol), andsodium acetate (1.052 g, 12.82 mmol) in dry dimethyl sulfoxide (4 mL)was degassed with argon. 1,1′-Bis-(diphenylphosphino)ferrocenepalladium(II) dichloride (0.156 g, 0.214 mmol) was added and the mixturewas heated at 90° C. for 3 h. After cooling down had taken place, thereaction mixture was partitioned between ethyl acetate (50 mL) and water(50 mL). The layers were separated and the organic layer was washed withwater (50 mL), brine (50 mL), dried with sodium sulfate and concentratedto give a red/brown crude product. The material was purified by flashcolumn chromatography (heptane:ethyl acetate) (10:0 to 8:2) affording acolorless oil (0.98 g, 65.3%).

¹H NMR (400 MHz, CDCl₃) δ ppm 5.53 (d, J=8 Hz, 1H), 5.44 (d, J=10 Hz,1H), 5.53 (t, J=7.2 Hz, 1H), 4.90 (s1H), 4.37 (d, J=5.8 Hz, 2H), 1.39(s, 9H), 1.25 (s, 12H).

Preparation of methyl8-[4-[(tert-butoxycarbonylamino)methyl]-3-fluoro-phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate

The general Suzuki coupling method described above was used to couplemethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate withtert-butylN-[[2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl]carbamate.Purification by flash silica column chromatography yielded methyl8-[4-[(tert-butoxycarbonylamino)methyl]-3-fluoro-phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylate(24 mg, 31.7%).

LC-MS: t=2.22 min (method 1); 481 (M+H)⁺.

Preparation of Compound 4 Preparation of8-[4-[(tert-butoxycarbonylamino)methyl]-3-fluoro-phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A solution of methyl8-(4-((tert-butoxycarbonylamino)methyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(24 mg, 0.050 mmol) and an aqueous 1N sodium hydroxide solution (0.500mL, 0.500 mmol) in MeOH (2 mL) was stirred at 50° C. for 2 h. Thereaction mixture was cooled. The MeOH was removed under reducedpressure, the residue was taken-up in water (5 mL) and then neutralizedwith a 1N hydrochloric acid solution (˜0.5 mL). A precipitation wasformed and the mixture was extracted with DCM (3×4 mL). The organiclayer was concentrated to give the acid (20 mg, 86%).

LC-MS: t=2.17 min (method 1); 467 (M+H)⁺; 465 (M−H)⁻.

Preparation of compound 48-[4-(aminomethyl)-3-fluoro-phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A solution of 4M hydrochloric acid in dioxane (1 mL, 4 mmol) was addedto a solution of8-(4-((tert-butoxycarbonylamino)methyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (24 mg, 0.051 mmol) in ACN (4 mL). The mixture was stirred for 4 hand a suspension was formed. The product compound 4 was collected byfiltration (17.7 mg, 78%).

LC-MS: t=2.54 min (method 2); 367 (M+H)⁺; 365 (M−H)⁻.

Preparation of Compound 5 Preparation of compound 58-(4-aminophenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

The general Suzuki coupling method described above was used to couplemethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate with4-butyloxycarbonyl-amino-phenyl boronic acid. Purification by flashsilica column chromatography yielded methyl8-[4-(tert-butoxycarbonylamino)-phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylateas a yellow solid. A solution of the obtained solid in a mixture of THF(1.5 mL) and aqueous 4N sodium hydroxide solution (0.5 mL) wasirradiated at 120° C. in a microwave oven for 10 min. The solvents wereevaporated and the residue taken in DCM (10 mL). The organic phase waswashed with water (10 mL), dried over sodium sulfate and concentrated todryness. A solution of the residue and TFA (0.5 mL) in DCM (0.5 mL) wasstirred at room temperature for 1.5 h. After evaporation of thesolvents, the product was purified by preparative HPLC yielding compound58-(4-aminophenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid (28.9 mg, 43.7%).

LC-MS: t=5.99 min (method 3); 349 (M+H)⁺; 332 (M-NH₃+H)⁺.

Preparation of Compound 6 Preparation of8-(4-aminophenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylicacid

The general Suzuki coupling method described above was used to coupleethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewith 4-butyloxy-carbonylamino-phenyl boronic acid. Purification by flashsilica column chromatography yielded ethyl8-[4-(tert-butoxycarbonylamino)-phenyl]-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylateas a yellow solid.

A solution of the obtained yellow solid in a mixture of THF (1.5 mL) andof aqueous 4N sodium hydroxide solution (0.5 mL) was irradiated at 120°C. in a microwave oven for 10 min. The solvents were evaporated and theresidue taken in DCM (10 mL). The organic phase was washed with water(10 mL), dried over sodium sulfate and concentrated to dryness. Asolution of the residue and TFA (0.5 mL) in DCM (0.5 mL) was stirred atroom temperature for 1.5 h. After evaporation of the solvents, theproduct was purified by preparative HPLC yielding compound 68-(4-aminophenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylicacid (16.4 mg, 52%).

LC-MS: t=6.01 min (method 3); 367 (M+H)⁺; 350 (M-NH₃+H)⁺; ¹H NMR (400MHz, DMSO-d6) δ ppm 13.89 (s, 1H), 9.38 (d, J=5.6 Hz, 1H), 8.24 (s, 1H),7.68 (d, J=8 Hz, 2H), 7.54 (d, J=8 Hz, 1H), 4.15 (s, 2H), 2.77 (s, 3H),2.48-2.52 (m, 1H), 1.03-1.05 (m, 2H), 0.78-0.80 (m, 2H).

Preparation of Compound 17 Preparation of ethyl8-[4-(tert-butoxycarbonylamino)-3-fluoro-phenyl]-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylate

The general Suzuki coupling method described above was used to coupleethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewith 3-fluoro-4-(tert-butyloxycarbonylamino-phenyl boronic acid.Purification by flash silica column chromatography yielded ethyl8-[4-(tert-butoxycarbonylamino)-3-fluoro-phenyl]-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylateas a yellow solid (280 mg, 61%).

LC-MS: t=2.85 min (method 1); 499 (M+H)⁺.

Preparation of compound 178-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-(tert-butoxycarbonylamino)-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(260 mg, 0.52 mmol) in a 4N hydrochloric acid solution (20 mL). Themixture was stirred for 20 min at 50° C. The product was lyophilized anddissolved in a mixture of THF (1 mL) and of an aqueous 4N sodiumhydroxide solution (1.31 mL, 5.2 mmol) before being irradiated at 140°C. in a microwave oven for 10 min. The product was purified bypreparative HPLC yielding compound 178-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3-carboxylicacid (5.1 mg, 2.5%).

LC-MS: t=2.24 min (method 1); 371 (M+H)⁺.

Preparation of the Compounds 1-11, 13, 15 and 17-89 from Scaffold a, B,C, D and E

Chemical names and structures of the compounds are given in Table 1.

The preparation of the compounds from scaffold A, C, D and E isdescribed below.

Compounds 1, 2, 3, 4, 5, 6, and 17 were also resynthesized according tothis method. Preparation of the corresponding potassium salts is alsodescribed. Compounds are then referred as such with the letter Kfollowing their number. If a compound is obtained as another salt (forexample hydrochloric salt), the compound is referred as such followedwith the appropriate suffix (for example HCl).

Most of the examples were prepared according to the general proceduresA-D described below. The preparation of the other examples is otherwisereported specifically in the experimental section. A number of boronateswere specifically prepared to be reacted with the scaffolds by generalprocedures A or A′. These boronate were made from commercialbromo-nitriles with general procedures E, F and G described below.

General Procedure A:

The quinolizine scaffold (1 eq.), boronate (1.3 eq.) and cesiumcarbonate (3 eq.) were added to a 3:1 mixture of 1,2-dimethoxyethane andwater (4 mL). The mixture was degassed with argon.1,1′-Bis-diphenylphosphine ferrocene palladium(II) dichloride (0.1 eq.)was added and the mixture was heated at 90° C. under an argon atmospherefor 1 h. The reaction mixture was allowed to cool down.

The usual work up procedure was as follows. The mixture was diluted withDCM (3 mL) and water was added (3 mL). The layers were separated using aphase separator and the aqueous layer was extracted with DCM (2×5 mL).The combined organic layers were dried over sodium sulfate andconcentrated in vacuum. An alternative work up procedure consisted infiltering the residue and rinsing it with DCM (5 mL) prior toconcentration of the solvents.

The crude product was purified by flash silica column chromatography anddried in vacuum to afford the desired product.

General Procedure A′ for Microwave Oven:

The quinolizine scaffold (1 eq.), boronate (1.3 eq.) and cesiumcarbonate (3 eq.) were added to a 3:1 mixture of 1,2-dimethoxyethane andwater (4 mL). The mixture was degassed with argon.1,1′-Bis-diphenylphosphine ferrocene palladium(II) dichloride (0.1 eq.)was added and the mixture was heated in a microwave oven at 150° C.under an argon atmosphere for 5 min. The reaction mixture was allowed tocool down.

The usual work up procedure was as follows. The mixture was diluted withDCM (3 mL) and water was added (3 mL). The layers were separated using aphase separator and the aqueous layer was extracted with DCM (2×5 mL).The combined organic layers were dried over sodium sulfate andconcentrated in vacuum. An alternative work up procedure consisted infiltering the residue and rinsing it with DCM (5 mL) prior toconcentration of the solvents.

The crude product was purified by flash silica column chromatography anddried in vacuum to afford the desired product.

General Procedure B:

The ester intermediate (1 eq.) was added to a solution of lithiumhydroxide (2 eq.) in a 1:1 mixture of THF and water. The reactionmixture was stirred for 18 h at 30° C. The mixture was acidified using1M HCl in water. The precipitate was filtered off and dried in vacuum toafford the desired product.

General Procedure C:

The BOC-protected amine intermediate (1 eq.) was suspended in DCM and 1M HCl in diethyl ether (20 eq.) was added. The reaction mixture wasstirred for 16 h. The precipitate was filtered off and dried in vacuumto afford the desired product.

General Procedure D:

The free acid (1 eq.) was added to a solution of potassium hydroxide(1.1 eq.) in water and stirred for 1 h. The solution was lyophilized toobtain the desired product

General Procedure E:

A suspension of the bromo-nitrile (1 eq.) in dry (8 ml) was cooled in anice bath under a N₂ atmosphere. Di-tert-butyl dicarbonate (2-3 eq.) wasadded followed by the careful addition of borane-THF complex (1M, 2.4eq.). The mixture was stirred at room temperature for 2 hours. Ifneeded, the borane-THF complex (1M, 2.4 eq.) was added again and themixture was stirred at room temperature overnight.

The mixture was quenched at 0° C. with water (8 ml). The layers wereseparated. The water layer was extracted with DCM. The combined organiclayers were dried over Na₂SO₄ and concentrated in vacuum to afford thecrude BOC protected amine.

General Procedure F:

To a solution of the bromo-amine (1eq.) in THF (8 ml) was added Et₃N (3eq.), 4-dimethylaminopyridine (0.1 eq.) and di-tert-butyl dicarbonate(2.2 eq.). Gas evolution was observed. The mixture was stirred at roomtemperature overnight. Water was added. The mixture was extracted withDCM. The combined organic layers were dried over Na₂SO₄ and concentratedin vacuum to afford the crude BOC protected bromo-amine.

General Procedure G:

A mixture of the bromide (1 eq.), bis(pinacolato)diboron (1.5 eq.) andpotassium acetate (3 eq.) in 1,4-dioxane (6 ml) and water (2 ml) wasdegassed with argon. 1,1′-bis-(diphenylphosphino)-ferrocene) palladiumdichloride (0.1 eq.) was added, the reaction tube was capped and heatedat 90° C. for 3 hours. Water was added and the mixture was extractedwith DCM. The combined organic layers were dried over Na₂SO₄ andconcentrated in vacuum to afford the crude pinacol ester.

Preparation and Characterization of Compounds 1-11, 13, 15 and 17-89Preparation of Compound 1 Preparation of methyl8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (200mg, 0.68 mmol) and2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (195 mg,0.82 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(191 mg, 76%).

ESI-MS m/z: 367 (M+H)⁺.

Preparation of compound 18-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(191 mg, 0.52 mmol) to afford the title compound 1 as a yellow solid(135 mg, 73%). ESI-MS m/z: 353 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm:14.05 (br s, 1H), 9.55 (d, J=7.3 Hz, 1H), 8.21 (s, 1H), 7.57 (d, J=7.3Hz, 1H), 7.16-7.34 (m, 2H), 6.92 (t, J=8.8 Hz, 1H), 5.74 (s, 2H), 2.89(s, 3H), 2.50-2.70 (m, 1H), 1.02-1.05 (m, 2H), 0.73-0.80 (m, 2H).

Preparation of compound 1K, potassium salt of compound 1—potassium8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (compound 1) (260 mg, 0.74 mmol) to afford the title compound Ksalt of compound 1 as a yellow solid (288 mg, 98%).

ESI-MS m/z: 353 (M−K+H)⁺.

Preparation of Compound 2 Preparation of2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline

Potassium acetate (7.08 g, 72 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (7.94 g, 31mmol) and 4-bromo-2,5-difluoro-aniline (5 g, 24 mmol) were dissolved in1,2-dimethoxyethane (60 mL), followed by the addition of1′-Bis-diphenylphosphine ferrocene palladium(II) dichloride (2.1 g, 2.6mmol). The reaction mixture was heated at 90° C. for 16 h. The reactionmixture was evaporated to dryness in vacuum. The mixture was coated ontohydromatrix and purified by flash silica column chromatography(heptane:ethyl acetate, 3:1). The product was dried in vacuum andcrystallized from DCM and heptane to give the title compound as a whitesolid (3.5 g, 57%).

ESI-MS m/z: 256.2 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm: 7.30 (dd, J=5.0Hz and J=11.1 Hz, 1H), 6.41 (q, J=6.8 Hz, 1H), 4.06 (br s, 1H), 1.33 (s,12H).

Preparation of methyl8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (500mg, 1.71 mmol) and2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline(667 mg, 2.06 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(500 mg, 75%).

ESI-MS m/z: 407 (M+Na)⁺.

Preparation of compound 28-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(1 g, 2.6 mmol) to afford the title compound 2 as a yellow solid (639mg, 66%).

ESI-MS m/z: 371 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.97 (s, 1H),9.32 (d, J=5.5 Hz, 1H), 8.24 (s, 1H), 7.21 (d, J=12.1 Hz, 1H), 7.03 (d,J=8.1 Hz, 1H), 6.92 (t, J=8.7 Hz, 1H), 6.00 (s, 2H), 2.85 (s, 3H),2.40-2.60 (m, 1H), 1.06-1.08 (m, 2H), 0.77-0.79 (m, 2H).

Preparation of compound 2K (potassium salt of compound 2) potassium8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Compound 2K—potassium salt of compound 2—Potassium8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (290 mg, 0.78 mmol) to afford the K salt of compound 2 as a yellowsolid (287 mg, 90%).

ESI-MS m/z: 371 (M−K+H)⁺.

Preparation of Compound 3 Preparation of ethyl8-(4-amino-3,5-dichloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.400 mmol),2,6-dichloro-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxa-borolan-2-yl)-aniline(138 mg, 0.48 mmol), (Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190mg in 0.8 mL of water, 1.80 mmol) in THF (5 mL) was degassed 3 timesunder N₂ and heated to 80° C. overnight. The reaction mixture wasdiluted with ethyl acetate (50 mL) and washed with water (2×20 mL). Theorganic phase was separated, dried, and concentrated. The residue waspurified by flash silica column chromatography (hexane:ethyl acetate,2:3) to afford the title compound as a yellow solid (140 mg, 81%).

ESI-MS m/z: 431 (M+H)⁺.

Preparation of compound 38-(4-amino-3,5-dichloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-amino-3,5-dichloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(140 mg, 0.326 mmol) in THF (6 mL) and water (2 mL) was treated withLiOH (55 mg, 1.31 mmol). The reaction was heated to 60° C. overnight andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (50 mL) and washed with brine (2×20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 3 as yellow solid (90 mg, 69%).

ESI-MS m/z: 403 (M+H)⁺; ¹H NMR (300 MHz, DMSO-d6) δ ppm: 9.30 (d, J=7.5Hz, 1H), 8.25 (s, 1H), 7.64 (d, J=7.5 Hz, 1H), 7.57 (s, 2H), 6.12 (s,2H), 2.92 (s, 3H), 2.54-2.51 (m, 1H), 1.13-1.10 (m, 2H), 0.84-0.83 (m,2H).

Preparation of Compound 4 Preparation of tert-butyl4-bromo-2-fluorobenzyl-carbamate

To a solution of (4-bromo-2-fluorophenyl) methanamine (2.5 g, 12 mmol)in DCM (50 mL) at 0° C. was added di-tert-butyl dicarbonate (4.01 g, 18mmol) and triethylamine (2.6 mL, 18 mmol). The mixture was allowed towarm to room temperature and stirred for 2 h. Water was added (50 mL)and the layers were separated. The organic layer was washed with brine,dried over sodium sulfate and concentrated in vacuum. The crude productwas purified by flash silica column chromatography (heptane:ethylacetate) (1:0 to 4:1) to afford the title compound as a colorless oil.(2.7 g, 71%).

¹H NMR (400 MHz, DMSO-d6) δ ppm 7.21-7.26 (m, 3H), 4.90 (br s, H),4.27-4.33 (m, 2H), 1.44 (s, 9H).

Preparation of tert-butyl2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzylcarbamate

A mixture of tert-butyl 4-bromo-2-fluorobenzyl-carbamate (1.3 g, 4.3mmol), bis(pinacolato)-diboron (1.6 g, 6.4 mmol), and sodium acetate(1.1 g, 13 mmol) in DMSO (dry) (4 mL) was degassed with argon.1,1′-Bis(diphenylphosphino)-ferrocene palladium(II) dichloride (0.156 g,0.214 mmol) was added and the mixture was heated at 90° C. for 3 h.After cooling, the reaction mixture was partitioned between ethylacetate (50 mL) and water (50 mL). The layers were separated and theorganic layer was washed with water (50 mL), brine, dried over sodiumsulfate and concentrated to give a red/brown crude product. The materialwas purified by flash silica column chromatography (heptane:ethylacetate) (1:0 to 4:1) to afford the title compound as a colorless oil(980 mg, 65%).

¹H NMR (400 MHz, DMSO-d6) δ ppm 7.53 (d, J=7.6 Hz, 1H), 7.45 (d, J=10.4Hz, 1H), 7.31-7.35 (m, 1H), 4.91 (br s, H), 4.36-4.40 (m, 2H), 1.44 (s,9H), 1.34 (s, 12H).

Preparation of methyl8-(4-((tert-butoxycarbonylamino)-methyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (46mg, 0.158 mmol) was dissolved in toluene (500 μL), ethanol (96%) (237μL) and aqueous sodium carbonate 2M (237 μL, 0.473 mmol). tert-Butyl2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamate(97 mg, 0.21 mmol) was added. The mixture was degassed with argon and1,1′-bis(diphenylphosphino)-ferrocene palladium(II) dichloride (11.5 mg,0.016 mmol) was added. The mixture was heated at 90° C. for 4 h. Aftercooling, the mixture was diluted with DCM (3 mL) and water (3 mL). Thelayers were separated and the aqueous layer was extracted with DCM (3×2mL). The organic layer was concentrated in vacuum. The crude product waspurified with flash silica column chromatography (heptane:ethyl acetate)(1:1 to 1:2) to afford the title compound as a yellow solid (24 mg,32%).

ESI-MS m/z: 481 (M+H)⁺.

Preparation of 8-(4-((tert-butoxycarbonylamino)methyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

Methyl 8-(4-((tert-butoxycarbonylamino)methyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(24 mg, 0.050 mmol) was dissolved in MeOH (2 mL) and 1M aqueous sodiumhydroxide (0.5 mL, 0.5 mmol) was added. The mixture was stirred at 50°C. for 2 h. After cooling, the MeOH was removed in vacuum and theresidue was dissolved in water (5 mL) and neutralized with 1M HCl (˜0.5mL). The precipitate formed was extracted with DCM (3×4 mL). The organiclayer was concentrated to afford the title compound as a yellow solid(20 mg, 86%).

ESI-MS m/z: 467 (M+H)⁺.

Preparation of compound 4HCl—HCl salt of compound4—8-(4-(aminomethyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

8-(4-((tert-Butoxycarbonylamino)methyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (24 mg, 0.051 mmol) was dissolved in ACN (4 mL). HCl 4M in dioxane(1 mL, 4 mmol) was added. The mixture was stirred for 4 h and asuspension was formed affording after filtration the title compound 4HClas a yellow solid (17.7 mg, 86%).

ESI-MS m/z: 453 (M+H)⁺.

Preparation of compound 4K—potassium salt of compound 4—potassium8-(4-(aminomethyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Compound 4K Potassium8-(4-(aminomethyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-(aminomethyl)-3-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (65 mg, 0.16 mmol) to afford the K salt of compound 4as a yellow solid (79 mg, 100%).

ESI-MS m/z: 367.0 (M−K+H)⁺.

Preparation of Compound 5 Preparation of methyl8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (500mg, 1.71 mmol) and(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-boronic acid (516 mg,2.06 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(850 mg, 100%).

ESI-MS m/z: 485 (M+Na)⁺, 463 (M+H)⁺.

Preparation of8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(((tert-Butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(1050 mg, 2.3 mmol) to afford the title compound as a yellow solid (950mg, 93%).

ESI-MS m/z: 449 (M+H)⁺.

Preparation of compound 5HCl—HCl salt of compound5—8-(4-(aminomethyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloric salt

8-(4-(Aminomethyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure C from8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (540 mg, 1.20 mmol) to afford the hydrochloric salt of the titlecompound 5 as a yellow solid (360 mg, 86%).

ESI-MS m/z: 349 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.07 (s, 1H),9.35 (d, J=7.3 Hz, 1H), 8.64 (s, 2H), 8.27 (s, 1H), 7.73 (d, J=7.8 Hz,2H), 7.56-7.63 (m, 3H), 4.13 (s, 2H), 2.86 (s, 3H), 2.40-2.60 (m, 1H),1.07-1.09 (m, 2H), 0.78-0.80 (m, 2H).

Preparation of compound 5K (potassium salt of compound 5) potassium8-(4-(amino-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-(amino-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-(amino-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloric salt (200 mg, 0.52 mmol) to afford the K salt ofcompound 5 as a yellow solid (153 mg, 76%).

ESI-MS m/z: 349 (M−K+H)⁺.

Preparation of Compound 6 Preparation of ethyl8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.31 mmol) and(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-boronic acid (101 mg,0.40 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(167 mg, 100%).

ESI-MS m/z: 495 (M+Na)⁺.

Preparation of8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(((tert-Butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(167 mg, 0.34 mmol) to afford the title compound as a yellow solid (150mg, 95%).

ESI-MS m/z: 467 (M+H)⁺.

Preparation of compound 6HCl (hydrochloric salt of compound 6)8-(4-amino-methyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloric salt

8-(4-Amino-methyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure C from8-(4-(((tert-butoxycarbonyl)-amino)-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (150 mg, 0.32 mmol) to afford the hydrochloric salt of the titlecompound 6 as a yellow solid (137 mg, 100%).

ESI-MS m/z: 367 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.9 (br s,1H), 9.41 (d, J=7.3 Hz, 1H), 8.50 (br s, 2H), 8.28 (s, 1H), 7.72 (d,J=7.8 Hz, 2H), 7.56 (d, J=7.8 Hz 2H), 4.16 (s, 2H), 2.79 (s, 3H),2.40-2.60 (m, 1H), 1.05-1.09 (m, 2H), 0.79-0.82 (m, 2H).

Preparation of compound 6K (potassium salt of compound 6) potassium8-(4-amino-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-amino-methyl)-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-aminomethyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloric salt (65 mg, 0.16 mmol) to afford the K salt ofcompound 6 as a yellow solid (79 mg, 100%).

ESI-MS m/z: 367 (M−K+H))⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.9 (br s,1H), 9.40 (d, J=7.3 Hz, 1H), 8.43 (br s, 2H), 8.28 (s, 1H), 7.72 (d,J=7.8 Hz, 2H), 7.56 (d, J=7.8 Hz, 2H), 4.16 (s, 2H), 2.79 (s, 3H),2.50-2.55 (m, 1H), 1.05-1.09 (m, 2H), 0.79-0.82 (m, 2H).

Preparation of Compound 7 Preparation of ethyl8-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(500 mg, 1.54 mmol) and4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (439 mg, 2.00mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded the title compound as a yellow solid (550 mg,93%).

ESI-MS m/z: 381 (M+H)⁺.

Preparation of compound 78-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(550 mg, 1.45 mmol) to afford the title compound 7 as a yellow solid(435 mg, 85%).

ESI-MS m/z: 353 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.95-14.02 (m,1H), 9.32 (d, J=5.8 Hz, 1H), 8.21 (s, 1H), 7.21 (d, J=6.8 Hz, 2H), 6.79(d, J=8.3 Hz, 2H), 2.85 (s, 3H), 2.50-2.60 (m, 1H), 1.05-1.07 (m, 2H),0.76-0.78 (m, 2H).

Preparation of compound 7K (potassium salt of compound 7) potassium8-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (460 mg, 0.74 mmol) to afford the title compound K salt of compound7 as a yellow solid (510 mg, 100%).

ESI-MS m/z: 353 (M−K+H)⁺.

Preparation of Compound 8 Preparation of methyl8-(4-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (150mg, 0.51 mmol) and4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (135 mg, 0.62mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded the title compound as a yellow solid (25 mg, 14%).

ESI-MS m/z: 349 (M+H)⁺.

Preparation of compound 88-(4-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(25 mg, 0.07 mmol) to afford the title compound compound 8 as a yellowsolid (18 mg, 77%).

ESI-MS m/z: 335 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.11 (s, 1H),9.24 (d, J=7.1 Hz, 1H), 8.22 (s, 1H), 7.53 (s, 1H), 7.30 (d, J=8.1 Hz,2H), 6.72 (d, J=8.4 Hz, 2H), 5.67 (s, 2H), 2.89 (s, 3H), 2.40-2.50 (m,1H), 1.05-1.07 (m, 2H), 0.73-0.75 (m, 2H).

Preparation of Compound 9 Preparation of methyl8-(3-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A using methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (195mg, 0.66 mmol) and (3-aminophenyl)-boronic acid (119 mg, 0.87 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded the title compound as a yellow solid (91 mg, 39%).

ESI-MS m/z: 349 (M+H)⁺.

Preparation of compound 98-(3-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-amino-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(91 mg, 0.26 mmol) to afford the title compound compound 9 as a yellowsolid (62.4 mg, 72%).

ESI-MS m/z: 335 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.20 (br s,1H), 9.32 (d, J=7.3 Hz, 1H), 8.26 (s, 1H), 7.52 (d, J=7.3 Hz, 1H),7.35-7.53 (m, 1H), 6.82-6.93 (m, 3H), 2.86 (s, 3H), 2.50-2.60 (m, 1H),1.00-1.10 (m, 2H), 0.75-0.85 (m, 2H).

Preparation of Compound 10 Preparation of methyl8-(3-S-((tert-butoxycarbonyl)-amino)-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol), tert-butyl-pyrrolidin-3-S-yl-carbamate (313 mg, 1.68mmol) and triethylamine (0.5 mL, 3.6 mmol) were added to 2-propanol (20mL). The reaction mixture was heated at 130° C. and stirred for 6 h. Thereaction mixture was cooled and evaporated to dryness. The crude productwas purified by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) and dried in vacuum to afford the title compound as a yellow foam(70 mg, 46%).

ESI-MS m/z: 442 (M+H)⁺.

Preparation of compound 108-(3-S-amino-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-S-amino-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-S-((tert-butoxycarbonyl)-amino)-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(76 mg, 0.17 mmol) to afford a yellow residue. The residue was treatedaccording to General Procedure C and purified using preparative LCMS toafford the title compound 10 as a yellow solid (5.5 mg, 10% over 2steps).

ESI-MS m/z: 328 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.03 (d, J=10.8Hz, 1H), 7.90 (s, 1H), 5.41 (s, 1H), 3.90-4.00 (m, 1H), 3.80-3.90 (m,1H), 3.65-3.80 (m, 1H), 3.50-3.60 (m, 1H), 3.30-3.40 (m, 2H), 2.58 (s,3H), 2.40-2.60 (m, 1H), 2.26-2.30 (m, 1H), 2.01-2.06 (m, 1H), 1.72-1.76(m, 1H), 0.95-0.98 (m, 2H), 0.57-0.60 (m, 2H).

Preparation of Compound 11 Preparation of ethyl8-(4-amino-2-chloro-5-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.4 mmol),3-chloro-6-methyl-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxa-borolan-2-yl)-aniline(128 mg, 0.48 mmol), (Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190mg in 0.8 mL of water, 1.80 mmol) in THF (5 mL) was degassed 3 timesunder N₂ and heated to 80° C. overnight. The reaction mixture wasdiluted with ethyl acetate (20 mL), and washed with water (20 mL). Theorganic phase was separated, dried, and concentrated. The residue waspurified by flash silica column chromatography (hexane:ethyl acetate,1:3) to afford the title compound as a yellow solid (70 mg, 43%).

ESI-MS m/z: 411 (M+H)⁺.

Preparation of compound 118-(4-amino-2-chloro-5-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-amino-2-chloro-5-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(70 mg, 0.171 mmol) in THF (6 mL) and water (2 mL) was treated with LiOH(29 mg, 0.690 mmol). The reaction was heated to 60° C. overnight andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (50 mL) and washed with brine (2×20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 11 as yellow solid (51 mg, 78%).

ESI-MS m/z: 383 (M+H)⁺; 1H NMR (300 MHz, DMSO-d6) δ ppm: 9.28 (d, J=7.5Hz, 1H), 8.24 (s, 1H), 7.43 (d, J=7.5 Hz, 1H), 6.99 (s, 1H), 6.81 (s,1H), 5.55 (s, 2H), 2.75 (s, 3H), 2.52-2.54 (m, 1H), 2.07 (s, 3H),1.01-1.03 (m, 2H), 0.70-0.74 (m, 2H).

Preparation of Compound 13 Preparation of methyl8-(5-((tert-butoxycarbonylamino)methyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(5-((tert-butoxycarbonylamino)methyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 5-(BOC-aminomethyl)thiophene-2-boronic acid (106 mg,0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(100.3 mg, 72%).

ESI-MS m/z: 469 (M+H)⁺.

Preparation of8-(5-((tert-butoxycarbonylamino)methyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

The compound8-(5-((tert-butoxycarbonylamino)methyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(5-((tert-butoxycarbonylamino)methyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(76.2 mg, 0.19 mmol) to afford the compound as a yellow solid (54 mg,68%).

ESI-MS m/z: 455 (M+H)⁺, 453 (M−H)⁻.

Preparation of compound 13HCl (hydrochloric salt of compound 13)8-(5-(aminomethyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

The compound8-(5-(aminomethyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride was prepared according to General Procedure C from8-(5-((tert-butoxycarbonylamino)methyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (105.3 mg, 0.23 mmol) to afford compound 13HCl as an orange solid(77.2 mg, 85%).

ESI-MS m/z: 355 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.27 (d, J=7.6Hz, 1H), 8.80 (s, 2H), 8.26 (s, 1H), 7.63 (d, J=7.3 Hz, 1H), 7.61 (d,J=3.8 Hz, 1H), 7.45 (d, J=3.8 Hz, 1H), 4.35 (s, 2H), 3.03 (s, 3H),2.50-2.57 (m, 1H), 1.06-1.11 (m, 2H), 0.73-0.77 (m, 2H).

Preparation of compound 13K (potassium salt of compound 13) potassium8-(5-(aminomethyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

The compound potassium8-(5-(aminomethyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(5-(aminomethyl)thiophen-2-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (77.2 mg, 0.20 mmol) to afford compound 13K as anorange solid (86.0 mg, 100%).

ESI-MS m/z: 355 (M−K+H)⁺, 353 (M−K−H)⁻.

Preparation of Compound 15 Preparation of ethyl8-(4-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.400 mmol), 4-methyl-phenyl boronic acid (65 mg, 0.48 mmol),(Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190 mg in 0.8 mL of water,1.80 mmol) in THF (5 mL) was degassed 3 times under N₂ and heated to 80°C. overnight. The reaction mixture was diluted with ethyl acetate (50mL), and washed with water (2×20 mL). The organic phase was separated,dried, and concentrated. The residue was purified by flash silica columnchromatography (hexane:ethyl acetate, 1:2) to afford the title compoundas a yellow solid (90 mg, 62%).

ESI-MS m/z: 362 (M+H)⁺.

Preparation of compound 158-(4-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-methylphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(90 mg, 0.249 mmol) in THF (6 mL) and water (2 mL) was treated with LiOH(43 mg, 1.02 mmol). The reaction was heated to 60° C. for 2 h andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (20 mL) and washed with brine (20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 15 as a yellow solid (65 mg, 78%).

ESI-MS m/z: 334 (M+H)⁺; ¹H NMR (300 MHz, DMSO-d6) δ ppm: 9.31 (d, J=7.2Hz, 1H), 8.24 (s, 1H), 7.54 (d, J=7.2 Hz, 1H), 7.44 (d, J=7.2 Hz, 2H),7.38 (d, J=7.2 Hz, 2H), 2.84 (s, 3H), 2.52-2.54 (m, 1H), 2.39 (s, 3H),1.03-1.06 (m, 2H), 0.75-0.77 (m, 2H).

Preparation of Compound 17 Preparation of ethyl8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(149 mg, 0.46 mmol) and2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (134mg, 0.58 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(18 mg, 10%).

ESI-MS m/z: 399 (M+H)⁺.

Preparation of compound 178-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(24 mg, 0.06 mmol) to afford the title compound 17 as a yellow solid (19mg, 85%).

ESI-MS m/z: 371 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.99 (br s,1H), 9.32 (s, 1H), 8.22 (s, 1H), 6.80-7.30 (m, 3H), 5.73 (s, 2H), 2.84(s, 3H), 2.50-2.60 (m, 1H), 1.04-1.08 (m, 2H), 0.76-0.80 (m, 2H).

Preparation of compound 17K (potassium salt of compound 17) potassium8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-amino-3-fluorop-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-amino-3-fluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (250 mg, 0.68 mmol) and potassium hydroxide to afford the titlecompound K salt of compound 17 as a yellow solid (276 mg, 90%).

ESI-MS m/z: 371 (M−K+H)⁺.

Preparation of Compound 18 Preparation of ethyl8-(3-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(3-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(102.5 mg, 0.32 mmol) and (3-amino-phenyl)-boronic acid (54.8 mg, 0.40mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded the title compound as a yellow solid (30 mg, 25%).

ESI-MS m/z: 381 (M+H)⁺.

Preparation of compound 188-(3-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(3-amino-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(140 mg, 0.37 mmol) to afford the title compound 18 as a yellow solid(30 mg, 25%).

ESI-MS m/z: 353 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.35 (d, J=5.8Hz, 1H), 8.25 (s, 1H), 7.22 (t, J=7.6 Hz, 1H), 6.71 (dd, J=1.6 Hz andJ=8.1 Hz, 1H), 6.57 (s, 1H), 6.51 (d, J=7.6 Hz, 1H), 5.40 (br s, 2H),2.82 (s, 3H), 2.53-2.63 (m, 1H), 1.04-1.07 (m, 2H), 0.76-0.78 (m, 2H).

Preparation of Compound 19 Preparation of ethyl8-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(500 mg, 1.54 mmol) and (4-carbamoyl-phenyl)-boronic acid (305.7 mg,1.85 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(540 mg, 85%).

ESI-MS m/z: 409 (M+H)⁺, 453 (M+HCOO)⁻.

Preparation of compound 198-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(540 mg, 1.32 mmol) and reacted for 10 days at 30° C. to afford thetitle compound 19 as a yellow soli: (360 mg, 71%).

ESI-MS m/z: 381 (M+H)⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm: 13.94 (s, 1H),9.42 (d, J=5.6 Hz, 1H), 8.28 (s, 1H), 8.16 (s, 1H), 8.08 (d, J=8.1 Hz,2H), 7.57-7.60 (m, 3H), 2.80 (s, 3H), 2.50-2.60 (m, 1H), 1.06-1.08 (m,2H), 0.76-0.80 (m, 2H).

Preparation of compound 19K (potassium salt of compound 19) potassium8-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-carbamoyl-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (310 mg, 0.82 mmol) to afford the title compound K salt of compound19 as a yellow solid (345 mg, 100%).

ESI-MS m/z: 381 (M−K+H)⁺.

Preparation of Compound 20 Preparation of methyl8-(4-amino-2-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-2-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (122mg, 0.51 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 94:6) afforded the title compound 20 as a yellowsolid (87 mg, 64%).

ESI-MS m/z: 367 (M+H)⁺.

Preparation of compound 208-(4-amino-2-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-2-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-2-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(87 mg, 0.24 mmol) to afford the title compound 20 as a yellow solid (50mg, 59%).

ESI-MS m/z: 353 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.27 (d, J=7.3Hz, 1H), 8.27 (s, 1H), 7.48 (s, 1H), 7.18 (t, J=8.6 Hz, 1H), 6.57 (dd,J=8.3 Hz, J=1.8 Hz, 1H), 6.49 (dd, J=13.4 Hz, J=1.8 Hz, 1H), 5.92 (s,2H), 2.80 (s, 3H), 2.40-2.60 (m, 1H), 1.03-1.06 (m, 2H), 0.70-0.72 (m,2H).

Preparation of Compound 21 Preparation of methyl8-(3-amino-4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-amino-4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (122mg, 0.51 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 94:6) afforded the title compound as a yellow solid(128 mg, 98%).

ESI-MS m/z: 367 (M+H)⁺.

Preparation of compound 218-(3-amino-4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Amino-4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-amino-4-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(123 mg, 0.35 mmol) to afford the title compound 21 as a yellow solid(60 mg, 49%).

ESI-MS m/z: 353 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.31 (d, J=7.3Hz, 1H), 8.26 (s, 1H), 7.50 (d, J=7.3 Hz, 1H), 7.18 (dd, J=8.3 Hz,J=11.4 Hz, 1H), 6.89 (dd, J=2.0 Hz, J=8.6 Hz, 1H), 6.63-6.67 (m, 1H),5.44 (s, 2H), 2.85 (s, 3H), 2.40-2.54 (m, 1H), 1.04-1.07 (m, 2H),0.76-0.78 (m, 2H).

Preparation of Compound 22 Preparation of methyl8-(3-amino-5-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-amino-5-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 3-amino-5-fluoro-phenyl-boronic acid (80 mg, 0.51mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 94:6) afforded the title compound as a yellow solid (108 mg,81%).

ESI-MS m/z: 367 (M+H)⁺.

Preparation of compound 228-(3-amino-5-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Amino-5-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-amino-5-fluorophenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(108 mg, 0.29 mmol) to afford the title compound 22 as a yellow solid(69 mg, 67%).

ESI-MS m/z: 353 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.22 (d, J=7.3Hz, 1H), 8.48 (s, 1H), 7.19 (d, J=7.6 Hz, 1H), 6.36-6.44 (m, 3H), 6.68(s, 2H), 2.81 (s, 3H), 2.40-2.50 (m, 1H), 1.02-1.04 (m, 2H), 0.70-0.72(m, 2H).

Preparation of Compound 23 Preparation of methyl8-(3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 3-fluoro-phenyl-boronic acid (71 mg, 0.51 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to94:6) afforded the title compound as a yellow solid (96 mg, 77%).

ESI-MS m/z: 352 (M+H)⁺.

Preparation of compound 238-(3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl1-cyclopropyl-8-(3-fluoro-phenyl)-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(96 mg, 0.27 mmol) to afford the title compound compound 23 as a yellowsolid (21 mg, 23%).

ESI-MS m/z: 338 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.34 (d, J=7.3Hz, 1H), 8.28 (s, 1H), 7.57-7.65 (m, 2H), 7.39-7.47 (m, 3H), 2.86 (s,3H), 2.40-2.50 (m, 1H), 1.07-1.09 (m, 2H), 0.80-0.82 (m, 2H).

Preparation of Compound 24 Preparation of methyl8-(4-amino-3-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-3-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (76mg, 0.26 mmol) and2-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (78 mg,0.31 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 94:6) afforded the title compound 24 as a yellowsolid (63 mg, 62%).

ESI-MS m/z: 387 (M+H)⁺.

Preparation of compound 248-(4-amino-3-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-3-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-3-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(63 mg, 0.16 mmol) to afford the title compound 24 as a yellow solid (37mg, 62%).

ESI-MS m/z: 369 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.21 (d, J=7.3Hz, 1H), 8.38 (s, 1H), 7.43 (s, 1H), 7.30 (s, 1H), 7.26 (d, J=14.5 Hz,1H), 6.94 (d, J=8.3 Hz, 1H), 5.82 (s, 2H), 2.85 (s, 3H), 2.40-2.50 (m,1H), 1.04-1.06 (m, 2H), 0.72-0.74 (m, 2H).

Preparation of Compound 25 Preparation of methyl8-(4-amino-3-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-3-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (76mg, 0.26 mmol) and2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (78mg, 0.31 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 94:6) afforded the title compound as a yellow solid(41 mg, 40%).

ESI-MS m/z: 379 (M+H)⁺.

Preparation of compound 258-(4-amino-3-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-3-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-3-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(41 mg, 0.11 mmol) to afford the title compound 25 as a yellow solid (29mg, 92%).

ESI-MS m/z: 365 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.27 (d, J=7.3Hz, 1H), 8.19 (s, 1H), 7.43 (d, J=7.3 Hz, 1H), 7.03 (s, 1H), 6.99 (dd,J=1.8 Hz, J=8.1 Hz, 1H), 6.80 (d, J=8.1 Hz, 1H), 5.38 (s, 2H), 3.86 (s,3H), 2.92 (s, 3H), 2.40-2.54 (m, 1H), 1.06-1.10 (m, 2H), 0.76-0.80 (m,2H).

Preparation of Compound 26 Preparation of methyl8-(4-acetamido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-acetamido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (77mg, 0.26 mmol) and (4-acetamido-phenyl)-boronic acid (56 mg, 0.31 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to94:6) afforded the title compound 26 as a yellow solid (93 mg, 90%).

ESI-MS m/z: 391 (M+H)⁺.

Preparation of compound 268-(4-acetamido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Acetamido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure: using methyl8-(4-acetamido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(93 mg, 0.24 mmol) to afford the title compound 26 as a yellow solid (45mg, 50%).

ESI-MS m/z: 377 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 10.20 (s, 1H),9.32 (d, J=7.3 Hz, 1H), 8.25 (s, 1H), 7.79 (d, J=8.4 Hz, 2H), 7.58 (d,J=7.3 Hz, 1H), 7.53 (d, J=8.1 Hz, 2H), 2.88 (s, 3H), 2.40-2.54 (m, 1H),2.10 (s, 3H), 1.07-1.09 (m, 2H), 0.77-0.79 (m, 2H).

Preparation of Compound 27 Preparation of methyl8-(4-(methylsulfonamido)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(methylsulfonamido)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (74mg, 0.25 mmol) andN-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-phenyl)-methane-sulfonamide(96 mg, 0.32 mmol). Compound precipitated from solution and was filteredoff and dried in vacuum to afford the title compound 27 as a yellowsolid (83 mg, 77%).

ESI-MS m/z: 427 (M+H)⁺.

Preparation of compound 278-(4-(methylsulfonamido)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(Methylsulfonamido)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-(methylsulfonamido)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(83 mg, 0.20 mmol) to afford the title compound 27 as a yellow solid (53mg, 64%).

ESI-MS m/z: 413 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 10.1 (s, 1H),9.33 (d, J=7.3 Hz, 1H), 8.26 (s, 1H), 7.55-7.58 (m, 3H), 7.39 (d, J=8.6Hz, 2H), 3.11 (s, 3H), 2.88 (s, 3H), 2.40-2.55 (m, 1H), 1.06-1.09 (m,2H), 0.78-0.80 (m, 2H).

Preparation of Compound 28 Preparation of methyl8-(4-(methylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(methylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (74mg, 0.25 mmol) andN-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (71 mg,0.31 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 94:6) afforded the title compound as a yellow solid(81 mg, 81%).

ESI-MS m/z: 363 (M+H)⁺.

Preparation of compound 288-(4-(methylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(Methylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-(methylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(81 mg, 0.22 mmol) to afford the title compound 28 as a yellow solid (55mg, 72%).

ESI-MS m/z: 349 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) 5 ppm: 9.27 (d, J=7.3Hz, 1H), 8.20 (s, 1H), 9.59 (d, J=7.3 Hz, 1H), 7.39 (d, J=8.6 Hz, 2H),6.70 (d, J=8.6 Hz, 2H), 6.28 (d, J=5.1 Hz, 1H), 2.90 (s, 3H), 2.76 (d,J=4.8 Hz, 3H), 2.40-2.52 (m, 1H), 1.06-1.08 (m, 2H), 0.75-0.77 (m, 2H).

Preparation of Compound 29 Preparation of methyl8-(pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (500mg, 1.71 mmol) and pyridin-4-yl-boronic acid (253 mg, 2.05 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded the title compound as a yellow solid (400 mg, 69%).

ESI-MS m/z: 373 (M+K)⁺, 357 (M+Na)⁺, 335 (M+H)⁺.

Preparation of compound 298-(pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(Pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl1-cyclopropyl-9-methyl-4-oxo-8-(pyridin-4-yl)-4H-quinolizine-3-carboxylate(400 mg, 1.20 mmol) to afford the title compound 29 as a yellow solid(262 mg, 68%).

ESI-MS m/z: 321 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.05 (s, 1H),9.36 (d, J=7.3 Hz, 1H), 8.79 (d, J=5.8 Hz, 2H), 8.30 (s, 1H), 7.55-7.59(m, 3H), 2.83 s, (3H), 2.50-2.60 (m, 1H), 1.05-1.10 (m, 2H), 0.79-0.84(m, 2H).

Preparation of compound 29K (potassium salt of compound 29) potassium8-(pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (compound 16) (250 mg, 0.78 mmol) to afford the K salt of titlecompound 29 as a yellow solid (290 mg, 100%).

ESI-MS m/z: 321 (M−K+H)⁺; 1H NMR (400 MHz, CD₃OD) δ ppm: 9.43 (d, J=7.3Hz, 1H), 8.72 (d, J=5.8 Hz, 2H), 8.62 (s, 1H), 7.58 (d, J=5.8 Hz, 2H),7.20 (d, J=3.6 Hz, 2H), 2.87 (s, 3H), 2.46-2.53 (m, 1H), 0.80-1.10 (m,2H), 0.84-0.86 (m, 2H).

Preparation of Compound 30 Preparation of methyl8-(pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and pyridin-3-ylboronic acid (63 mg, 0.51 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to94:6) afforded the title compound as a yellow solid (100 mg, 87%).ESI-MS m/z: 335 (M+H)⁺.

Preparation of compound 308-(pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(Pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl1-cyclopropyl-9-methyl-4-oxo-8-(pyridin-3-yl)-4H-quinolizine-3-carboxylate(100 mg, 0.30 mmol) to afford the title compound 30 as a yellow solid(84 mg, 87%).

ESI-MS m/z: 321 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.35 (d, J=7.3Hz, 1H), 8.73-8.78 (s, 2H), 8.28 (s, 1H), 8.02-8.05 (m, 1H), 7.61-7.65(m, 2H), 2.86 (s, 3H), 2.40-2.54 (m, 1H), 1.06-1.10 (m, 2H), 0.80-0.83(m, 2H).

Preparation of Compound 31 Preparation of2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline

Potassium acetate (317 mg, 3.22 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (300 mg,1.18 mmol) and 4-bromo-2-methyl-aniline (200 mg, 1.08 mmol) weredissolved in DMSO (3 mL). The reaction mixture was degassed using argon.1,1′-Bis-diphenylphosphine ferrocene palladium(II) dichloride (24 mg,0.03 mmol) was added and the reaction mixture was heated at 80° C. for 5h. The reaction mixture was cooled to room temperature and diluted withethyl acetate (20 mL). The organic layer was washed with saturatedsodium bicarbonate (20 mL) and brine (20 mL). The organic layer wasdried with sodium sulfate, filtered and purified by flash silica columnchromatography (heptane:ethyl acetate, 0-40%) to obtain a crude mixtureof starting material and product. The mixture was purified usingreversed phase column chromatography to obtain the title compound as aclear oil (50 mg, 21%).

ESI-MS m/z: 234 (M+H)⁺.

Preparation of methyl8-(4-amino-3-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-3-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (50mg, 0.17 mmol) and4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline (52 mg, 0.22mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded the title compound as a yellow solid (31 mg, 50%).

ESI-MS m/z: 363 (M+H)⁺.

Preparation of compound 318-(4-amino-3-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-3-methy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-3-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(31 mg, 0.09 mmol) to afford the title compound 31 as a yellow solid (19mg, 60%).

ESI-MS m/z: 349 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.99 (s, 1H),9.16-9.25 (m, 1H), 8.21-8.34 (m, 1H), 7.52-7.54 (m, 2H), 7.16-7.20 (m,1H), 6.76 (d, J=8.1 Hz, 1H), 5.43 (s, 2H), 2.99 (s, 3H), 2.89 (s, 3H),2.40-2.50 (m, 1H), 1.03-1.07 (m, 2H), 0.70-0.75 (m, 2H).

Preparation of Compound 32 Preparation of methyl8-(2-fluoro-pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(2-fluoro-pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 2-fluoro-pyridin-4-yl-boronic acid (72 mg, 0.51mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 94:6) afforded the title compound as a yellow solid (107 mg,89%).

ESI-MS m/z: 353 (M+H)⁺.

Preparation of compound 328-(2-fluoro-pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(2-Fluoro-pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(2-fluoro-pyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(107 mg, 0.30 mmol) to afford the title compound 32 as a yellow solid(53 mg, 52%).

ESI-MS m/z: 339 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.21-9.31 (m,1H), 8.52 (s, 1H), 8.39-8.47 (m, 1H), 7.20-7.60 (m, 3H), 2.80 (s, 3H),2.50-2.60 (m, 1H), 0.98-1.13 (m, 2H), 0.71-0.81 (m, 2H).

Preparation of Compound 33 Preparation of methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-pyridin-2-amine (113 mg,0.51 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 94:6) afforded the title compound as a yellow solid(135 mg, 100%).

ESI-MS m/z: 350 (M+H)⁺.

Preparation of compound 338-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-Amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(61 mg, 0.17 mmol) to afford the title compound 33 as a yellow solid (34mg, 60%).

ESI-MS m/z: 336 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.20 (d, J=7.6Hz, 1H), 8.47 (s, 1H), 8.33 (s, 1H), 8.12 (s, 1H), 7.60 (dd, J=2.3 Hz,J=8.6 Hz, 1H), 8.29 (d, J=6.6 Hz, 1H), 6.59 (d, J=8.8 Hz, 1H), 6.39 (s,2H), 2.84 (s, 3H), 2.50-2.60 (m, 1H), 1.23 (s, 2H), 1.03-1.07 (m, 4H)0.70-0.72 (m, 2H).

Preparation of compound 33K (potassium salt of compound 33) potassium8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (510 mg, 1.52 mmol) to afford the K salt of compound 33 as a yellowsolid (564 mg, 99%).

ESI-MS m/z: 336 (M−K+H)⁺.

Preparation of Compound 34 Preparation of methyl1-cyclopropyl-8-(1H-indol-5-yl)-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-indol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 1H-indol-5-yl-boronic acid (83 mg, 0.51 mmol) toafford the title compound as a yellow solid (60 mg, 47%).

ESI-MS m/z: 373 (M+H)⁺.

Preparation of compound 348-(1H-indol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-indol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(60 mg, 0.16 mmol) to afford the title compound compound 34 as a yellowsolid (20 mg, 35%).

ESI-MS m/z: 359 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 11.35 (s, 1H),9.24 (d, J=7.0 Hz, 1H), 8.48 (s, 1H), 7.70 (s, 1H), 7.50-7.60 (m, 2H),7.46 (s, 1H), 7.31 (d, J=7.6 Hz, 1H), 7.22 (d, J=7.8 Hz, 1H), 6.54 (s,1H), 2.84 (s, 3H), 2.50-2.60 (m, 1H), 1.00-1.08 (m, 2H), 0.72-0.76 (m,2H).

Preparation of Compound 35 Preparation of methyl8-(1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol), cesium carbonate (335 mg, 1.03 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (92 mg, 0.38mmol) were added to a mixture of 1,2-dimethoxyethane (3 mL) and water (1mL). The mixture was degassed with argon.1,1′-Bis-(diphenylphosphino)-ferrocene) palladium dichloride (28 mg,0.03 mmol) was added. The reaction mixture was heated at 150° C. in amicrowave oven under argon atmosphere for 0.25 h. The reaction mixturewas cooled. The mixture was diluted with DCM (3 mL) and water was added(3 mL). The layers were separated using a phase separator and theaqueous layer was extracted with DCM (2×5 mL). The combined organiclayers were concentrated in vacuum. The crude product was purified byflash silica column chromatography (DCM:MeOH) (1:0 to 9:1) and dried invacuum to afford the title compound as a yellow solid (93 mg, 72%).

ESI-MS m/z: 374 (M+H)⁺.

Preparation of compound 358-(1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(555 mg, 1.49 mmol) to afford the title compound compound 35 as a yellowsolid (200 mg, 37%).

ESI-MS m/z: 360 (M+H)⁺; ([M−H]⁻=358.2; 100%); 1H NMR (400 MHz, DMSO-d6)δ ppm: 14.13 (s, 1H), 13.35 (s, 1H), 9.36 (d, J=7.3 Hz, 1H), 8.25 (d,J=15.9 Hz, 2H), 8.01 (s, 1H), 7.74 (d, J=8.6 Hz, 1H), 7.67 (d, J=7.6 Hz,1H), 7.54 (d, J=9.6 Hz, 1H), 2.90 (s, 3H), 2.40-2.60 (m, 1H), 1.07-1.11(m, 2H), 0.79-0.82 (m, 2H).

Preparation of compound 35K (potassium salt of compound 35) potassium8-(1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (180 mg, 0.50 mmol) to afford the K salt of compound 35 as a yellowsolid (195 mg, 98%).

ESI-MS m/z: 360 (M−K+H)⁺.

Preparation of Compound 36 Preparation of methyl8-(4-ureido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-ureido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (75mg, 0.26 mmol) and1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-phenyl)-urea (84 mg,0.32 mmol). Compound precipitated from solution and was filtered off anddried in vacuum to afford the title compound as a yellow solid (29 mg,26%).

ESI-MS m/z: 392 (M+H)⁺.

Preparation of compound 368-(4-ureido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Ureido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-ureido-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(28 mg, 0.07 mmol) to afford the title compound compound 36 as a yellowsolid (15 mg, 58%).

ESI-MS m/z: 378 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.10 (s, 1H),9.30 (d, J=7.6 Hz, 1H), 9.09 (s, 1H), 8.23 (s, 1H), 7.57-7.63 (m, 3H),7.46 (d, J=8.6 Hz, 2H), 6.04 (s, 2H), 2.89 (s, 3H), 2.40-2.55 (m, 1H),1.06-1.08 (m, 2H), 0.76-0.78 (m, 2H).

Preparation of Compound 37 Preparation of methyl8-(4-(dimethylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(dimethylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (77mg, 0.26 mmol) and (4-(dimethylamino)-phenyl)-boronic acid (56 mg, 0.34mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 94:6) afforded the title compound as a yellow solid (39 mg,31%).

ESI-MS m/z: 377 (M+H)⁺.

Preparation of compound 378-(4-(dimethylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(Dimethylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-(dimethylamino)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(34 mg, 0.09 mmol) to afford the title compound 37 as a yellow solid(5.1 mg, 15%).

ESI-MS m/z: 363 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.10-9.20 (m,1H), 8.44 (s, 1H), 7.20-7.40 (m, 3H), 6.80-6.85 (m, 2H), 2.88 (s, 6H),2.82 (s, 3H), 2.50-2.60 (m, 1H), 0.98-1.05 (m, 1H), 0.65-0.70 (m, 1H).

Preparation of Compound 38 Preparation of methyl8-(3-S-((tert-butoxycarbonyl)-amino)-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol), tert-butyl-pyrrolidin-3-S-yl-carbamate (313 mg, 1.68mmol) and triethylamine (0.5 mL, 3.6 mmol) were added to 2-propanol (20mL). The reaction mixture was heated at 130° C. and stirred for 6 h. Thereaction mixture was cooled and evaporated to dryness. The crude productwas purified by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) and dried in vacuum to afford the title compound as a yellow foam(70 mg, 46%).

ESI-MS m/z: 442 (M+H)⁺.

Preparation of compound 38HCl (hydrochloric salt of compound 38)8-(3-S-amino-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-S-amino-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-S-((tert-butoxycarbonyl)-amino)-pyrrolidin-1-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(76 mg, 0.17 mmol) to afford a yellow residue. The residue was treatedaccording to General Procedure C and purified using preparative LCMS toafford the hydrochloric salt of the title compound 38 as a yellow solid(5.5 mg, 10% over 2 steps).

ESI-MS m/z: 328 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.03 (d, J=10.8Hz, 1H), 7.90 (s, 1H), 5.41 (s, 1H), 3.90-4.00 (m, 1H), 3.80-3.90 (m,1H), 3.65-3.80 (m, 1H), 3.50-3.60 (m, 1H), 3.30-3.40 (m, 2H), 2.58 (s,3H), 2.40-2.60 (m, 1H), 2.26-2.30 (m, 1H), 2.01-2.06 (m, 1H), 1.72-1.76(m, 1H), 0.95-0.98 (m, 2H), 0.57-0.60 (m, 2H).

Preparation of Compound 39 Preparation of compound 398-piperazin-1-yl-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A mixture of methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (200mg, 0.69 mmol), N-(tert-butoxycarbonyl)-piperazine (256 mg, 1.38 mmol)and NaHCO₃ (259 mg, 14.64 mmol) in ACN (8.6 mL) was heated in amicrowave at 120° C. for 20 min. DMF (2 mL) was added to the mixture andthe reaction was heated in a microwave at 120° C. for 20 min. Then thereaction was heated in a microwave at 130° C. for 30 min twice. Thereaction mixture was diluted with ethyl acetate (30 mL), and washed withwater (30 mL). After extraction of the aqueous phase with ethyl acetate(2×30 mL), the organic phases were combined washed with brine, driedwith magnesium sulfate, filtrated, and concentrated. The residue wasdissolved in TFA (1 mL) and agitated for 1 hour prior to evaporation ofthe solvent. The residue was dissolved in THF (1 mL) and an aqueous 4MNaOH solution (0.79 mL) and heated in a microwave at 120° C. for 10 min.More aqueous 4M NaOH solution (0.5 mL) was added and the reaction washeated in a microwave at 120° C. for 10 min. The mixture was evaporatedand the residue purified by preparative HPLC. The title compoundcompound 39 was obtained after lyophilization (28 mg, 12.5%).

High-Res MS: calculated 328.1656 (M+H)⁺. found 328.1642 (M+H)⁺.

Preparation of Compound 40 Preparation of compound 408-[(3S)-3-amino-1-piperidyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylicacid

A mixture of methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (300mg, 1 mmol), 3-S—N-(tert-butoxycarbonyl)-amino)-piperidine (413 mg, 2mmol) and NaHCO₃ (390 mg, 14.64 mmol) in ACN (13 mL) was heated in amicrowave at 120° C. for 20 min, and twice at 130° C. for 30 min. Thereaction mixture was diluted with ethyl acetate (30 mL), and washed withwater (30 mL). After extraction of the aqueous phase with ethyl acetate(2×30 mL), the organic phases were combined washed with brine, driedwith magnesium sulfate, filtrated, and concentrated. The residue wasdissolved in TFA (1 mL) and agitated for 1 hour prior to evaporation ofthe solvent. The residue was dissolved in THF (1 mL) and an aqueous 4MNaOH solution (1.4 mL) and heated in a microwave at 100° C. for 10 minand at 120° C. for 10 min. The mixture was evaporated and the residuepurified by preparative HPLC. The title compound 40 was obtained afterlyophilization (65 mg, 19%).

High-Res MS: calculated m/z 342.1812 (M+H)⁺. found m/z 342.1787 (M+H)⁺.

Preparation of Compound 41 Preparation of methyl8-(4-cyano-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-cyano-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (123mg, 0.42 mmol) and 4-cyano-phenyl-boronic acid (132 mg, 0.90 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded the title compound as a yellow solid (142 mg, 94%).

ESI-MS m/z: 359 (M+H)⁺.

Preparation of compound 418-(4-carbamoyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Carbamoyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-cyano-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(123 mg, 0.35 mmol) and purified by preparative LCMS to afford the titlecompound 41 as a yellow solid (9.1 mg, 7%).

ESI-MS m/z: 363 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) 5 ppm: 9.25 (d, J=7.3Hz, 1H), 8.24 (s, 1H), 8.12 (s, 1H), 8.04 (d, J=8.1 Hz, 1H), 7.61 (d,J=8.1 Hz, 1H), 7.50 (s, 1H), 7.35 (d, J=7.1 Hz, 1H), 2.81 (s, 3H),2.50-2.60 (m, 1H), 1.02-1.10 (m, 2H), 0.71-0.79 (m, 2H).

Preparation of Compound 42 Preparation of compound 428-(4-carboxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Carboxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-cyano-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(123 mg, 0.35 mmol). Preparative LC-MS purification afforded the titlecompound 42 as a yellow solid (9.3 mg, 7%).

ESI-MS m/z: 364 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) 5 ppm: 9.36 (d, J=7.3Hz, 1H), 8.50 (br s, 1H), 8.08 (d, J=8.1 Hz, 1H), 7.47 (d, J=8.1 Hz,1H), 7.32 (br s, 1H), 2.86 (s, 3H), 2.46-2.55 (m, 1H), 1.02-1.12 (m,1H), 0.78-0.82 (m, 1H).

Preparation of Compound 43 Preparation of ethyl8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.32 mmol),2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline(667 mg, 2.06 mmol), tricyclohexylphosphine (105 mg, 0.41 mmol) andcesium fluoride (482 mg, 3.2 mmol) were added to ACN (5 mL). Thereaction mixture was degassed with argon. Palladium(II) acetate (24 mg,0.11 mmol) was added. The reaction mixture was heated at 85° C. for 2 h.The reaction mixture was cooled and evaporated in vacuum. Purificationby flash silica column chromatography (DCM:MeOH) (1:0 to 9:1) affordedthe title compound as a yellow solid (100 mg, 35%).

ESI-MS m/z: 403 (M+H)⁺.

Preparation of compound 438-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-2,5-difluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-amino-2,5-difluoro-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.24 mmol). The compound was purified using preparative LCMSand dried in vacuum to afford the title compound 43 as a yellow solid(11 mg, 12%).

ESI-MS m/z: 389 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.0 (br s,1H), 9.30 (d, J=4.3 Hz, 1H), 8.26 (s, 1H), 7.25 (q, J=6.6 Hz, 1H), 6.72(t, J=7.6 Hz, 1H), 6.02 (s, 2H), 2.84 (s, 3H), 2.50-2.60 (m, 1H),0.98-1.12 (m, 2H), 0.63-0.83 (m, 2H).

Preparation of Compound 44 Preparation of2,6-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline

Potassium acetate (7.08 g, 72 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (6.7 g, 26mmol) and 4-bromo-2,6-difluoroaniline (5 g, 24 mmol) were dissolved inDMSO (30 mL), followed by the addition of 1′-Bis-diphenylphosphineferrocene palladium(II) dichloride (0.53 g, 0.7 mmol). The reactionmixture was heated at 80° C. for 16 h. The reaction mixture was dilutedin ethyl acetate (150 mL) and washed with saturated sodium bicarbonateand brine (2×100 mL). The organic layer was collected, dried over sodiumsulfate and dried in vacuum. Purification by flash silica columnchromatography (hexane:ethyl acetate, 3:2) afforded the title compoundas a white solid (4.5 g, 73%).

ESI-MS m/z: 256 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm: 7.20-7.30 (m,2H), 3.93 (br s, 2H), 1.32 (s, 12H).

Preparation of methyl8-(4-amino-3,6-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-amino-3,6-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (500mg, 1.71 mmol) and2,6-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-aniline(667 mg, 2.06 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(1140 mg, 86%).

ESI-MS m/z: 423 (M+K)⁺, 407 (M+Na)⁺, 385 (M+H)⁺.

Preparation of compound 448-(4-amino-3,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Amino-3,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-amino-3,6-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(1140 mg, 3.0 mmol) to afford the title compound 44 as a yellow solid(916 mg, 97%).

ESI-MS m/z: 371 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.08 (s, 1H),9.26 (d, J=7.3 Hz, 1H), 8.22 (s, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.25 (d,J=7.3 Hz, 2H), 5.80 (s, 2H), 2.89-2.91 (m, 3H), 2.40-2.60 (m, 1H),1.07-1.09 (m, 2H), 0.77-0.79 (m, 2H).

Preparation of compound 44K (potassium salt of compound 44) potassium8-(4-amino-3,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-amino-3,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-amino-3,5-difluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (300 mg, 0.81 mmol) to afford the K salt of compound 44 as a yellowsolid (335 mg, 100%).

ESI-MS m/z: 371 (M−K+H)⁺.

Preparation of Compound 45 Preparation of methyl8-(4-cyano-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-cyano-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 4-cyano-3-fluoro-phenyl-boronic acid (85 mg, 0.51mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 94:6) afforded the title compound as a yellow solid (135 mg,98%).

ESI-MS m/z: 377 (M+H)⁺.

Preparation of compound 458-(4-cyano-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Cyano-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from me methyl8-(4-cyano-3-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(135 mg, 0.36 mmol) to afford the title compound 45 as a yellow solid(37 mg, 28%).

ESI-MS m/z: 363 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) 5 ppm: 14.0 (br s,1H), 9.30 (d, J=7.3 Hz, 1H), 8.33 (s, 1H), 8.15 (t, J=7.6 Hz, 1H), 7.81(d, J=7.6 Hz, 1H), 7.59-7.61 (m, 1H), 7.38-7.52 (m, 1H), 2.82 (s, 3H),2.40-2.60 (m, 1H), 1.00-1.18 (m, 2H), 0.73-0.92 (m, 2H).

Preparation of Compound 46 Preparation of ethyl8-(4-cyano-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(4-cyano-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(151 mg, 0.47 mmol) and 4-cyano-phenyl-boronic acid (82 mg, 0.56 mmol).Purification by flash silica column chromatography (DCM:MeOH, 1:0 to9:1) afforded the title compound as a yellow solid (189 mg, 100%).

ESI-MS m/z: 391 (M+H)⁺.

Preparation of compound 468-(4-carboxy-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Carboxy-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-cyano-phenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(123 mg, 0.35 mmol). Preparative LC-MS purification afforded the titlecompound 46 as a yellow solid (12 mg, 7%).

ESI-MS m/z: 382 (M+H)⁺; 1H NMR (400 MHz, CD₃OD) δ ppm 9.40 (br s, 1H),8.50 (br s, 1H), 8.11 (d, J=8.1 Hz, 2H), 7.40 (d, J=8.1 Hz, 2H), 2.83(s, 3H), 2.43-2.55 (m, 1H), 0.99-1.15 (m, 2H), 0.82-0.86 (m, 2H).

Preparation of Compound 47 Preparation of methyl8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (500mg, 1.71 mmol) and tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(467.0 mg, 1.51 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as abrown oil (830 mg, 100%).

ESI-MS m/z: 461 (M+Na)⁺, 439 (M+H)⁺.

Preparation of8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1-(tert-Butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B using methyl8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(830 mg, 1.89 mmol) to afford the title compound as a brown oil (628 mg,78%).

ESI-MS m/z: 425 (M+H)⁺.

Preparation of compound 478-(1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1,2,3,6-Tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure C from8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid and purified by preparative LCMS to afford the title compound 47 asa yellow solid (164 mg, 47%).

ESI-MS m/z: 325 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) 5 ppm: 14.05 (s, 1H),9.32 (d, J=7.3 Hz, 1H), 8.25 (s, 1H), 7.41 (d, J=7.3 Hz, 1H), 5.91 (s,1H), 3.79 (s, 2H), 3.39-3.41 (m, 2H), 2.93 (s, 3H), 2.54-2.59 (s, 2H),2.40-2.60 (m, 1H), 1.03-1.11 (m, 2H), 0.73-0.77 (m, 2H).

Preparation of Compound 48 Preparation of methyl8-(1-(tert-butoxycarbonyl)-1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1-(tert-butoxycarbonyl)-1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and tert-butyl3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrole-1-carboxylate(151.0 mg, 0.51 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 94:6) afforded the title compound as ayellow solid (89 mg, 61%).

ESI-MS m/z: 423 (M+H)⁺.

Preparation of compound 488-(1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1-(tert-butoxycarbonyl)-1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(89 mg, 0.22 mmol) followed by BOC removal according to GeneralProcedure C. Purification by preparative LCMS afforded the titlecompound 48 as a yellow solid (22 mg, 7% over 2 steps).

ESI-MS m/z: 309 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm: 14.14 (s, 1H),11.58 (s, 1H), 9.18 (d, J=7.6 Hz, 1H), 8.14 (s, 1H), 7.78 (d, J=7.6 Hz,1H), 7.53 (s, 1H), 7.01 (s, 1H), 6.64 (s, 1H), 3.04 (s, 3H), 2.50-2.60(m, 1H), 1.03-1.12 (m, 2H), 0.68-0.72 (m, 2H).

Preparation of compound 48K (potassium salt of compound 48) potassium8-(1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

The title compound potassium8-(1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-pyrrol-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (34.0 mg, 0.11 mmol) to afford the K salt compound 48 as a yellowsolid (37.9 mg, 94%).

ESI-MS m/z: 309 (M−K+H)⁺, 307 (M+H)⁺. ([M−K-H].

Preparation of Compound 49 Preparation of ethyl8-(4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.400 mmol), 4-fluoro-phenyl boronic acid (67 mg, 0.48 mmol),(Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190 mg in 0.8 mL of water,1.80 mmol) in THF (5 mL) was degassed 3 times under N₂ and heated to 80°C. overnight. The reaction mixture was diluted with ethyl acetate (50mL), and washed with water (2×20 mL). The organic phase was separated,dried, and concentrated. The residue was purified by flash silica columnchromatography (hexane:ethyl acetate, 1:2) to afford the title compoundas a yellow solid (120 mg, 82%).

ESI-MS m/z: 366 (M+H)⁺.

Preparation of compound 498-(4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-fluoro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(120 mg, 0.329 mmol) in THF (6 mL) and water (2 mL) was treated withLiOH (55 mg, 1.31 mmol). The reaction was heated to 60° C. for 2 h andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (50 mL) and washed with brine (2×20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 49 as a yellow solid (110 mg, 99%).

ESI-MS m/z: 338 (M+H)⁺; ¹H NMR (300 MHz, DMSO-d6) δ ppm: 9.37 (d, J=7.5Hz, 1H), 8.30 (s, 1H), 7.70-7.61 (m, 3H), 7.51-7.45 (m, 2H), 2.89 (s,3H), 2.53-2.51 (m, 1H), 1.13-1.10 (m, 2H), 0.83-0.82 (m, 2H).

Preparation of Compound 50 Preparation of ethyl8-(4-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.400 mmol), 4-chloro-phenyl boronic acid (75 mg, 0.48 mmol),(Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190 mg in 0.8 mL of water,1.80 mmol) in THF (5 mL) was degassed 3 times under N₂ and heated to 80°C. overnight. The reaction mixture was diluted with ethyl acetate (50mL), and washed with water (2×20 mL). The organic phase was separated,dried, and concentrated. The residue was purified by flash silica columnchromatography (hexane:ethyl, acetate 2:3) to afford the title compoundas a yellow solid (95 mg, 62%).

ESI-MS m/z: 382 (M+H)⁺.

Preparation of compound 508-(4-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-chloro-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(95 mg, 0.249 mmol) in THF (6 mL) and water (2 mL) was treated with LiOH(43 mg, 1.02 mmol). The reaction was heated to 60° C. for 4 h andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (50 mL) and washed with brine (2×20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 50 as yellow solid (49 mg, 56%).

ESI-MS m/z: 354 (M+H)⁺; ¹H NMR (300 MHz, DMSO-d6) 5 ppm: 9.34 (d, J=7.2Hz, 1H), 8.27 (s, 1H), 7.56-7.68 (m, 5H), 2.84 (s, 3H), 2.52-2.55 (m,1H), 2.39 (s, 3H), 1.05-1.08 (m, 2H), 0.78-0.80 (m, 2H).

Preparation of Compound 51 Preparation of methyl8-(4-hydroxy-phenyl)-91-cyclopropyl-1-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-hydroxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (500mg, 1.71 mmol) and 4-hydroxy-phenyl-boronic acid (283 mg, 2.06 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded the title compound as a yellow solid (610 mg, 100%).

ESI-MS m/z: 388 (M+K)⁺, 372 (M+Na)⁺, 350 (M+H)⁺.

Preparation of compound 518-(4-hydroxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Hydroxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl1-cyclopropyl-8-(4-hydroxy-phenyl)-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(610 mg, 1.74 mmol) to afford the title compound 51 as a yellow solid(383 mg, 65%).

ESI-MS m/z: 336 (M+H)⁺, 334.20 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) 5 ppm:14.12 (s, 1H), 10.00 (s, 1H), 9.30 (d, J=7.3 Hz, 1H), 8.23 (s, 1H), 7.57(d, J=7.6 Hz, 1H), 7.43 (d, J=8.6 Hz, 2H), 6.96 (d, J=8.6 Hz, 2H), 2.88(s, 3H), 2.40-2.60 (m, 1H), 1.05-1.08 (m, 2H), 0.76-0.78 (m, 2H).

Preparation of compound 51K (potassium salt of compound 51) potassium8-(4-hydroxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-hydroxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from1-cyclopropyl-8-(4-hydroxy-phenyl)-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (370 mg, 0.81 mmol) to afford the K salt of compound 51 as a yellowsolid (429 mg, 100%).

ESI-MS m/z: 336 (M−K+H)⁺, 334 (M−K−H)⁻.

Preparation of Compound 52 Preparation of ethyl8-(4-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.400 mmol), 4-methoxy-phenyl boronic acid (73 mg, 0.48 mmol),(Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190 mg in 0.8 mL of water,1.80 mmol) in THF (5 mL) was degassed 3 times under N₂ and heated to 80°C. overnight. The reaction mixture was diluted with ethyl acetate, andwashed with water. The organic phase was separated, dried, andconcentrated. The residue was purified by flash silica columnchromatography (hexane:ethyl acetate, 1:2) to afford the title compoundas a yellow solid (68 mg, 45%).

ESI-MS m/z: 378 (M+H)⁺.

Preparation of compound 528-(4-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-methoxy-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(68 mg, 0.180 mmol) in THF (6 mL) and water (2 mL) was treated with LiOH(33 mg, 0.78 mmol). The reaction was heated to 60° C. overnight andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (50 mL) and washed with brine (2×20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 52 as yellow solid (22 mg, 35%).

ESI-MS m/z: 350 (M+H)⁺; ¹H NMR (300 MHz, DMSO-d6) δ ppm: 9.30 (d, J=7.5Hz, 1H), 8.23 (s, 1H), 7.57 (d, J=7.5 Hz, 1H), 7.52 (d, J=8.4 Hz, 2H),7.12 (d, J=7.5 Hz, 2H), 3.84 (s, 3H), 2.86 (s, 3H), 2.51-2.53 (m, 1H),1.04-1.06 (m, 2H), 0.75-0.78 (m, 2H).

Preparation of Compound 53 Preparation of ethyl8-(4-hydroxy-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

A mixture of ethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (120mg, 0.400 mmol), 4-hydroxy-methyl-phenyl boronic acid (73 mg, 0.48mmol), (Ph₃P)₂PdCl₂ (28 mg, 0.040 mmol), and Na₂CO₃ (190 mg in 0.8 mL ofwater, 1.80 mmol) in THF (5 mL) was degassed 3 times under N₂ and heatedto 80° C. overnight. The reaction mixture was diluted with ethyl acetate(50 mL) and washed with water (2×20 mL). The organic phase wasseparated, dried, and concentrated. The residue was purified by flashsilica column chromatography (hexane:ethyl acetate, 1:3 to pure ethylacetate) to afford the title compound as a yellow solid (60 mg, 40%).

ESI-MS m/z: 378 (M+H)⁺.

Preparation of compound 538-(4-hydroxy-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

A solution of ethyl8-(4-hydroxy-methyl-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(60 mg, 0.159 mmol) in THF (6 mL) and water (2 mL) was treated with LiOH(27 mg, 0.642 mmol). The reaction was heated to 60° C. overnight andacidified with 1N HCl to pH 4. The precipitate was dissolved with ethylacetate (50 mL) and washed with brine (2×20 mL). The organic phase wasseparated, dried, and concentrated. The precipitate was filtered toafford the title compound 53 as yellow solid (21 mg, 38%).

ESI-MS m/z: 350 (M+H)⁺; ¹H NMR (300 MHz, DMSO-d6) δ ppm: 9.38 (d, J=7.2Hz, 1H), 8.30 (s, 1H), 7.62 (d, J=7.5 Hz, 1H), 7.56-7.59 (m, 4H), 5.41(t, 1H), 4.64 (d, J=5.7 Hz, 2H), 2.91 (s, 3H), 2.51-2.54 (m, 1H),1.10-1.13 (m, 2H), 0.81-0.83 (m, 2H).

Preparation of Compound 54 Preparation of tert-butyl7-bromo-1-hydroxy-2-oxo-1,2,3,4-tetrahydroquinolin-3-yl-carbamate

To a suspension of3-amino-7-bromo-1-hydroxy-3,4-dihydroquinolin-2(1H)-one hydrochloride(203 mg, 0.692 mmol) in DCM (10 mL) was added di-tert-butyl dicarbonate(226 mg, 1.04 mmol) and triethylamine (0.482 mL, 3.46 mmol). Thesuspension was stirred at room temperature for 16 h. Water (20 mL) wasadded to the clear solution and the layers were separated. The organiclayer was washed with brine, dried over sodium sulfate and concentratedin vacuum. The crude product was purified by flash silica columnchromatography (heptane:ethyl acetate) (1:0 to 7:3) to afford the titlecompound as a white solid (102 mg, 41%).

ESI-MS m/z: 303, 301 (M−tBu+H)⁺.

Preparation of tert-butyl2-oxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-3-ylcarbamate

Sodium acetate (20.7 mg, 0.252 mmol) and bis(pinacolato)diboron (32.0mg, 0.126 mmol) were placed as solids in a flask under argon. tert-Butyl7-bromo-1-hydroxy-2-oxo-1,2,3,4-tetrahydroquinolin-3-ylcarbamate (30 mg,0.084 mmol) in DMSO (dry) (1 mL) was added and the mixture was degassedwith argon. trans-Bis(triphenylphosphine)-palladium(II) dichloride (5.9mg, 8.4 μmol) was added and the reaction mixture was heated at 40° C.for 2 h. After cooling the reaction mixture was concentrated andpurified by flash silica column chromatography (heptane:ethyl acetate)(95:5 to 3:2) to afford the title compound as a white solid (13.5 mg,41%).

¹H NMR (400 MHz, DMSO-d6) δ ppm 7.78 (s, 1H), 7.46 (d, J=7.6 Hz, 1H),7.21 (d, J=7.6 Hz, 1H), 5.60 (br s, 1H), (4.34 (br s, 1H), 3.48-3.53 (m,1H), 2.81-2.89 (m, 2H), 1.47 (s, 9H), 1.34 (s, 12H).

Preparation of methyl8-(3-(tert-butoxycarbonylamino)-2-oxo-1,2,3,4-tetrahydroquinolin-7-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (45mg, 0.15 mmol) was dissolved in toluene (330 μL), ethanol (96%) (243 μL)and 2M aqueous sodium carbonate solution (231 μL, 0.463 mmol).tert-Butyl2-oxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-3-ylcarbamate(78 mg, 0.20 mmol) was added and the mixture was degassed with argon.1,1′-Bis(diphenylphosphino)-ferrocene palladium(II) dichloride (11.3 mg,0.015 mmol) was added and the mixture was heated at 80° C. under anargon atmosphere for 16 h. After cooling, the mixture was diluted withDCM (3 mL) and water (3 mL) and the layers were separated. The aqueouslayer was extracted with DCM (3×2 mL). The combined organic layers wereconcentrated and the yellow crude product was purified with flash silicacolumn chromatography (heptane/ethyl acetate) (1:0 to 0:1) to afford thetitle compound as a yellow solid (43 mg, 53%).

ESI-MS m/z: 518 (M+H)⁺.

Preparation of8-(3-(tert-butoxycarbonylamino)-2-oxo-1,2,3,4-tetrahydroquinolin-7-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

Methyl8-(3-(tert-butoxycarbonylamino)-2-oxo-1,2,3,4-tetrahydroquinolin-7-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(43 mg, 0.083 mmol) was dissolved in MeOH (2 mL) and sodium hydroxide 1Msolution in water (0.5 mL, 0.5 mmol) was added. The mixture was stirredat 50° C. for 2 h. After cooling, the MeOH was removed in vacuum. Theresidue was dissolved in water (5 mL) and then neutralized with 1M HCl(˜0.5 mL). The precipitate formed was stirred at room temperatureovernight. The mixture was extracted with DCM (3×4 mL). The organiclayers were concentrated to afford the title compound as a yellow solid(29 mg, 69%).

ESI-MS m/z: 504 (M+H)⁺.

Preparation of compound 54HCl (hydrochloric salt of compound 54)8-(3-amino-2-oxo-1,2,3,4-tetrahydroquinolin-7-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

8-(3-(tert-Butoxycarbonylamino)-2-oxo-1,2,3,4-tetrahydroquinolin-7-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (29 mg, 0.046 mmol) was dissolved in ACN (4 mL). HCl 4M in dioxane(1 mL, 4 mmol) was added. The mixture was stirred for 4 h and asuspension was formed. The solvents were evaporated and the crude wastriturated with diethyl ether (4 mL) to afford the hydrochloric salt ofthe title compound 54 as a yellow solid (20 mg, 98%).

ESI-MS m/z: 404 (M+H)⁺.

Preparation of Compound 55 Preparation of methyl8-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-ol (90.8 mg,0.41 mmol). The precipitate was rinsed with DCM and dried in a vacuumstove to afford quantitatively the title compound as a yellow solid.

ESI-MS m/z: 351 (M+H)⁺.

Preparation of compound 558-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-Hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.34 mmol) to afford compound 55 as a yellow solid (113 mg, 99% in twosteps).

ESI-MS m/z: 337 (M+H)⁺, 335 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.08 (s, 1H), 12.19 (s, 1H), 9.26 (d, J=7.3 Hz, 1H), 8.25 (s, 1H), 7.77(s, 1H), 7.71 (d, J=9.4 Hz, 1H), 7.57 (s, 1H), 6.51 (d, J=9.6 Hz, 1H),2.88 (s, 3H), 2.40-2.60 (m, 1H), 1.05-1.07 (m, 2H), 0.76-0.77 (m, 2H).

Preparation of compound 55K (potassium salt of compound 55) potassium8-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-hydroxypyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (113.0 mg, 0.34 mmol) to afford the K salt of compound 55 as ayellow solid (123.0 mg, 95%).

ESI-MS m/z: 337 (M−K+H)⁺, 335 (M−K−H)⁻.

Preparation of Compound 56 Preparation of methyl8-(3-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (3-hydroxyphenyl)-boronic acid (56.6 mg, 0.41 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded quantitatively the title compound as a yellow solid.

ESI-MS m/z: 350 (M+H)⁺; 348 (M−H)⁻.

Preparation of compound 568-(3-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.34 mmol) to afford compound 56 as a yellow solid (81 mg, 71% in twosteps).

ESI-MS m/z: 336 (M+H)⁺; 334 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.10 (s, 1H), 9.83 (s, 1H), 9.32 (d, J=7.3 Hz, 1H), 8.27 (s, 1H), 7.53(d, J=7.3 Hz 1H), 7.38 (t, J=7.8 Hz, 1H), 6.89-6.94 (m, 3H), 2.86 (s,3H), 2.40-2.60 (m, 1H), 1.04-1.10 (m, 2H), 0.77-0.78 (m, 2H).

Preparation of compound 56K (potassium salt of compound 56) potassium1-cyclopropyl-8-(3-hydroxyphenyl)-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D8-(3-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (81.0 mg, 0.24 mmol) to afford the K salt of compound 56 as ayellow (94.0 mg, 100%).

ESI-MS m/z: 336 (M−K+H)⁺.

Preparation of Compound 57 Preparation of methyl8-(2-aminopyrimidin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(2-aminopyrimidin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (90.8mg, 0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(96 mg, 80%).

ESI-MS m/z: 351 (M+H)⁺; 349 (M−H)⁻.

Preparation of compound 578-(2-aminopyrimidin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(2-Aminopyrimidin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(2-aminopyrimidin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(96 mg, 0.27 mmol). Purification by preparative HPLC afforded compound57 as a yellow solid (10.8 mg, 11%).

ESI-MS m/z: 337 (M+H)⁺; 335 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.29(d, J=7.6 Hz, 1H), 8.54 (s, 2H), 8.50 (s, 1H), 8.23 (s, 1H), 7.64 (d,J=7.4 Hz, 1H), 7.22 (s, 2H), 2.91 (s, 3H), 2.40-2.60 (m, 1H), 1.07-1.09(m, 2H), 0.76-0.78 (m, 2H).

Preparation of Compound 58 Preparation of methyl8-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (3-fluoropyridin-4-yl)-boronic acid (65.3 mg, 0.41mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded the title compound as a yellow solid (78 mg, 65%).

ESI-MS m/z: 353 (M+H)⁺; 351 (M−H)⁻.

Preparation of compound 588-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(78 mg, 0.22 mmol) to afford compound 58 as a yellow solid (55 mg, 73%).

ESI-MS m/z: 339 (M+H)⁺; 337 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.03 (s, 1H), 9.36 (d, J=7.1 Hz, 1H), 8.84 (s, 1H), 8.67 (d, J=4.3 Hz,1H), 8.34 (s, 1H), 7.60-7.70 (m, 1H), 7.57 (d, J=5.8 Hz, 1H), 2.81 (s,3H), 2.50-2.60 (m, 1H), 1.06-1.08 (m, 2H), 0.78-0.80 (m, 2H).

Preparation of compound 58K (potassium salt of compound 58) potassium8-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(3-fluoropyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (55.0 mg, 0.16 mmol) to afford the K salt of compound 58 as ayellow solid (38.0 mg, 58%).

ESI-MS m/z: 339 (M−K+H)⁺; 337 (M−K−H)⁻.

Preparation of Compound 59 Preparation of ethyl8-(pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.34 mmol) and pyridin-4-ylboronic acid (52.1 mg, 0.42 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded quantitatively the title compound as a yellow solid.

ESI-MS m/z: 367 (M+H)⁺.

Preparation of compound 591-cyclopropyl-7-fluoro-9-methyl-4-oxo-8-(pyridin-4-yl)-4H-quinolizine-3-carboxylicacid

8-(Pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-8-(pyridin-4-yl)-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.34 mmol) to afford compound 59 as a yellow solid (75 mg, 72% in twosteps).

ESI-MS m/z: 339 (M+H)⁺; 1H NMR (400 MHz, DMSO) δ ppm 13.67 (s, 1H), 9.36(d, J=6.0 Hz, 1H), 8.81 (d, J=5.8 Hz, 2H), 8.31 (s, 1H), 7.50 (d, J=5.6Hz, 2H), 2.79 (s, 3H), 2.40-2.60 (m, 1H), 1.06-1.09 (m, 2H), 0.80-0.82(m, 2H).

Preparation of compound 59K (potassium salt of compound 59) potassium8-(pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(pyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (75.0 mg, 0.22 mmol) to afford the K salt of compound 59 as ayellow solid (49.8 mg, 58%).

ESI-MS m/z: 339 (M−K+H)⁺, 337 (M−K−H)⁻.

Preparation of Compound 60 Preparation of methyl8-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.34 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (93.7 mg,0.43 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded quantitatively the title compound as ayellow solid.

ESI-MS m/z: 382 (M+H)⁺.

Preparation of compound 608-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-Aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.32 mmol) to afford compound 60 as a yellow solid (81 mg, 72% in twosteps).

ESI-MS m/z: 354 (M+H)⁺, 352 (M−H)⁻; 1H NMR (400 MHz, DMSO) δ ppm 13.94(s, 1H), 9.34 (d, J=5.8 Hz, 1H), 8.24 (s, 1H), 8.12 (s, 1H), 7.68 (d,J=8.8 Hz, 1H), 7.10 (s, 2H), 6.79 (d, J=8.8 Hz, 1H), 2.87 (s, 3H),2.40-2.60 (m, 1H), 1.06-1.08 (m, 2H), 0.78-0.80 (m, 2H).

Preparation of compound 60K (potassium salt of compound 60) potassium8-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

potassium8-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-aminopyridin-3-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (81.0 mg, 0.23 mmol) to afford the K salt of compound 60 as ayellow solid (70.4 mg, 73%).

ESI-MS m/z: 354 (M−K+H)⁺, 352 (M−K−H)⁻.

Preparation of Compound 61 Preparation of ethyl8-(4-hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(4-hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.34 mmol) and (4-hydroxyphenyl)boronic acid (55.4 mg, 0.40mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded the title compound as a yellow solid (80 mg, 68%).

ESI-MS m/z: 382 (M+H)⁺.

Preparation of compound 618-(4-hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(4-hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(80 mg, 0.21 mmol) to afford compound 61 as a yellow solid (71 mg, 96%).

ESI-MS m/z: 354 (M+H)⁺; 1H NMR (400 MHz, DMSO) δ ppm 13.96 (s, 1H),10.04 (s, 1H), 9.34 (d, J=5.8 Hz, 1H), 8.23 (s, 1H), 7.31 (d, J=8.1 Hz,2H), 6.97 (d, J=8.1 Hz, 2H), 2.81 (s, 3H), 2.40-2.60 (m, 1H), 1.04-1.06(m, 2H), 0.77-0.79 (m, 2H).

Preparation of compound 61K (potassium salt of compound 61) potassium8-(4-hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium18-(4-Hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-Hydroxyphenyl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (71 mg, 0.20 mmol) to afford the K salt of compound 61 as a yellowsolid (56.4 mg, 67%).

ESI-MS m/z: 354 (M−K+H)⁺, 352 (M−K−H)⁻.

Preparation of Compound 62 Preparation of methyl8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(100 mg, 0.32 mmol) andtert-butyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(124.5 mg, 0.40 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded quantitatively the titlecompound as a yellow solid.

ESI-MS m/z: 471 (M+H)⁺.

Preparation of8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1-(tert-Butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.32 mmol) to afford the title compound as a yellow solid (98 mg, 69%in two steps).

ESI-MS m/z: 443 (M+H)⁺, 441 (M−H)⁻.

Preparation of compound 62HCl (hydrochloric salt of compound 62)8-(1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

The compound8-(1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride was prepared according to General Procedure C from8-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (98 mg, 0.22 mmol) to afford the HCl salt of compound 62 as ayellow solid (87 mg, 100%).

ESI-MS m/z: 343 (M+H)⁺, 341 (M−H)⁻; 1H NMR (400 MHz, DMSO) δ ppm 13.90(s, 1H), 9.33 (d, J=5.8 Hz, 1H), 9.28 (s, 1H), 9.25 (s, 1H), 5.96 (s,1H), 3.83 (s, 2H), 3.42 (s, 1H), 3.35-3.41 (m, 2H), 3.23 (s, 1H), 2.97(s, 3H), 2.50-2.58 (m, 1H), 1.04-1.07 (m, 2H), 0.77-0.79 (m, 2H).

Preparation of compound 62K (potassium salt of compound 62) potassium8-(1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (87 mg, 0.22 mmol) to afford the K salt of compound62 as a yellow solid (78.5 mg, 89%).

ESI-MS m/z: 343 (M−K+H)⁺.

Preparation of Compound 63 Preparation of methyl8-(4-(2,2,2-trifluoroacetyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(2,2,2-trifluoroacetyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and2,2,2-trifluoro-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethanone(123.3 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as ayellow solid (93 mg, 63%).

ESI-MS m/z: 448 (M+H₃O)⁺, 428 (M−H)⁻.

Preparation of compound 638-(4-(2,2,2-trifluoroacetyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylylicacid

8-(4-(2,2,2-Trifluoroacetyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylylicacid was prepared according to General Procedure B from methyl8-(4-(2,2,2-trifluoroacetyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(93 mg, 0.22 mmol). Purification by preparative HPLC afforded compound63 as a yellow solid (7.9 mg, 9%).

ESI-MS m/z: 434 (M+H₂O+H)⁺; 414 (M−H)⁻; 1H NMR (400 MHz, MeOD-d6) δ ppm9.42 (s, 1H), 8.44 (d, J=13.9 Hz, 1H), 7.82 (s, 2H), 7.61 (s, 1H), 7.50(s, 2H), 2.94 (s, 3H), 2.53 (s, 1H), 1.13 (s, 2H), 0.84 (s, 2H).

Preparation of Compound 64 Preparation of methyl8-(4-(acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (4-(acetamidomethyl)phenyl)-boronic acid (79.3 mg,0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(137 mg, 99%).

ESI-MS m/z: 405 (M+H)⁺.

Preparation of compound 648-(4-(acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(Acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-(acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(137 mg, 0.34 mmol) to afford compound 64 as a yellow solid (111.5 mg,84%).

ESI-MS m/z: 391 (M+H)⁺; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.33 (d, J=7.3Hz, 1H), 8.48 (t, J=5.8 Hz, 1H), 8.27 (s, 1H), 7.56 (s, 1H) 7.53 (d,J=8.1 Hz, 2H), 7.45 (d, J=8.1 Hz, 2H), 7.36 (d, J=5.8 Hz, 2H), 2.86 (s,3H), 2.40-2.60 (m, 1H), 1.91 (s, 3H), 1.06-1.08 (m, 2H), 0.78-0.80 (m,2H).

Preparation of compound 64K (potassium salt of compound 64) potassium8-(4-(Acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-(Acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-(acetamidomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (108.0 mg, 0.28 mmol) to afford the K salt of compound 64 as ayellow solid (111.5 mg, 93%).

ESI-MS m/z: 391 (M−K+H)⁺.

Preparation of Compound 65 Preparation of methyl8-(2-methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(2-methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (2-methylpyridin-4-yl)boronic acid (56.3 mg, 0.41mmol). Purification by flash silica column chromatography (DCM:MeOH)(1:0 to 9:1) afforded quantitatively the title compound as a yellowsolid.

ESI-MS m/z: 349 (M+H)⁺.

Preparation of compound 658-(2-methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(2-Methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(2-methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.34 mmol) to afford compound 65 as a yellow solid (71.7 mg, 63%).

ESI-MS m/z: 335 (M+H)⁺, 333 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.04 (s, 1H), 9.35 (d, J=7.3 Hz, 1H), 8.65 (d, J=5.0 Hz, 1H), 8.30 (s,1H), 7.54 (d, J=7.3 Hz, 1H), 7.44 (s, 1H), 7.36 (d, J=4.8 Hz, 1H), 2.84(s, 3H), 2.40-2.60 (m, 1H), 2.59 (s, 3H), 1.06-1.09 (m, 2H), 0.79-0.81(m, 2H).

Preparation of compound 65K (potassium salt of compound 65) potassium8-(2-methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(2-methylpyridin-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from1-cyclopropyl-9-methyl-8-(2-methylpyridin-4-yl)-4-oxo-4H-quinolizine-3-carboxylicacid (68.0 mg, 0.20 mmol) to afford the K salt of compound 65 as ayellow solid (71.7 mg, 94%).

ESI-MS m/z: 335 (M−K+H)⁺, 333 (M−K−H)⁻.

Preparation of Compound 66 Preparation of methyl8-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (102.7mg, 0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(101 mg, 78%).

ESI-MS m/z: 380 (M+H)⁺, 378 (M−H)⁻.

Preparation of compound 668-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-Hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(101 mg, 0.27 mmol) to afford compound 66 as a yellow solid (73.2 mg,75%).

ESI-MS m/z: 366 (M+H)⁺, 364 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm13.93 (s, 1H), 9.58 (s, 1H), 9.28 (d, J=7.0 Hz, 1H), 8.29 (s, 1H), 7.53(s, 1H), 7.11 (s, 1H), 6.96-6.99 (m, 2H), 3.85 (s, 3H), 2.89 (s, 3H),2.40-2.60 (m, 1H), 1.06-1.09 (m, 2H), 0.77-0.79 (m, 2H).

Preparation of compound 66K (potassium salt of compound 66) potassium8-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-hydroxy-3-methoxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (117.9 mg, 0.32 mmol) to afford the K salt of compound 66 as ayellow solid (73.2 mg, 53%).

ESI-MS m/z: 366 (M−K+H)⁺, 364 (M−K−H)⁻.

Preparation of Compound 67 Preparation of methyl8-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate(120.8 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as ayellow solid (54 mg, 49%).

ESI-MS m/z: 324 (M+H)⁺, 322 (M−H)⁻.

Preparation of compound 678-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(54 mg, 0.17 mmol) to afford compound 67 as a yellow solid (28 mg, 54%).

ESI-MS m/z: 310 (M+H)⁺, 308 (M−H)⁻; 1H NMR (400 MHz, MeOD-d6) δ ppm 9.34(d, J=7.6 Hz, 1H), 8.358 (s, 1H), 8.20 (s, 2H), 7.67 (d, J=7.3 Hz, 1H),3.11 (s, 3H), 2.49-2.53 (m, 1H), 1.12-1.15 (m, 2H), 0.76-0.79 (m, 2H).

Preparation of compound 67K (potassium salt of compound 67) potassium8-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-pyrazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (25.0 mg, 0.08 mmol) to afford the K salt of compound 67 as ayellow solid (27.2 mg, 88%).

ESI-MS m/z: 310 (M−K+H)⁺, 308 (M−K−H)⁻.

Preparation of Compound 68 Preparation of methyl8-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one (106.5mg, 0.41 mmol). The precipitate was rinsed with DCM and dried in avacuum stove to afford the title compound as a yellow solid (116 mg,87%).

ESI-MS m/z: 389 (M+H)⁺.

Preparation of compound 688-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(54 mg, 0.14 mmol) to afford compound 68 as a yellow solid (46 mg, 88%).

ESI-MS m/z: 375 (M+H)⁺, 373 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.04 (s, 1H), 9.36 (d, J=7.1 Hz, 1H), 8.77 (s, 1H), 8.30 (s, 1H), 7.86(d, J=7.8 Hz, 1H), 7.77 (s, 1H), 7.64 (d, J=7.6 Hz, 1H), 7.57 (d, J=6.6Hz, 1H), 4.50 (s, 2H), 2.86 (s, 3H), 2.40-2.60 (m, 1H), 1.06-1.09 (m,2H), 0.79-0.81 (m, 2H).

Preparation of compound 68K (potassium salt of compound 68) potassium8-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D8-(3-oxoisoindolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (44.0 mg, 0.12 mmol) to afford the K salt of compound 68 as ayellow solid (46.5 mg, 93%).

ESI-MS m/z: 375 (M−K+H)⁺, 373 (M−K−H)⁻.

Preparation of Compound 69 Preparation ofN,2-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

N,2-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline wasprepared according to General Procedure G with4-bromo-N,2-dimethylaniline and bis(pinacolato)diboron. Purification byflash silica column chromatography (heptane: ethyl acetate) (1:0 to 1:1)followed by recrystallization in DCM and heptane afforded the titlecompound as an off-white solid (104 mg, 17%).

ESI-MS m/z: 248 (M+H)⁺; 1H NMR (400 MHz, DMSO) δ ppm 7.63 (dd, J=8.1 Hz,J=1.0 Hz, 1H), 7.50 (s, 1H), 6.58 (d, J=8.1 Hz, 1H), 3.83 (br s, 1H),2.91 (s, 3H), 2.12 (s, 3H), 1.32 (s, 12H).

Preparation of methyl8-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) andN,2-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(101.5 mg, 0.41 mmol). The residue was rinsed with DCM and dried in avacuum stove to afford the title compound as a yellow solid (50 mg,39%).

ESI-MS m/z: 377 (M+H)⁺, 375 (M−H)⁻.

Preparation of compound 698-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(50 mg, 0.13 mmol) to afford compound 69 as a yellow solid (44.3 mg,92%).

ESI-MS m/z: 363 (M+H)⁺, 361 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.15 (s, 1H), 9.27 (d, J=7.3 Hz, 1H), 8.19 (s, 1H), 7.60 (d, J=7.3 Hz,1H), 7.34 (d, J=7.8 Hz, 1H), 7.26 (s, 1H), 6.64 (d, J=8.3 Hz, 1H),6.62-6.65 (m, 1H), 2.91 (s, 3H), 2.82 (d, J=4.8 Hz, 3H), 2.40-2.60 (m,1H), 2.17 (s, 3H), 1.05-1.10 (m, 2H), 0.74-0.78 (m, 2H).

Preparation of compound 69K (potassium salt of compound 69) potassium8-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(3-methyl-4-(methylamino)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (41.0 mg, 0.11 mmol) to afford the K salt of compound 69 as ayellow solid (44.3 mg, 97%).

ESI-MS m/z: 363 (M−K+H)⁺, 361 (M−K−H)⁻.

Preparation of Compound 70 Preparation of methyl8-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (3-fluoro-4-hydroxyphenyl)boronic acid (64.1 mg, 0.41mmol). The residue was rinsed with DCM and dried in a vacuum stove toafford the title compound as a yellow solid (95 mg, 76%).

ESI-MS m/z: 368 (M+H)⁺, 366 (M−H)⁻.

Preparation of compound 708-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(95 mg, 0.26 mmol) to afford compound 70 as a yellow solid (87.8 mg,96%).

ESI-MS m/z: 354 (M+H)⁺, 352 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.10 (s, 1H), 10.47 (s, 1H), 9.30 (d, J=7.3 Hz, 1H), 8.24 (s, 1H), 7.58(d, J=7.1 Hz, 1H), 7.46 (d, J=12.4 Hz, 1H), 7.24 (d, J=7.6 Hz, 1H), 7.14(t, J=8.7 Hz, 1H), 2.88 (s, 3H), 2.40-2.60 (m, 1H), 1.07-1.09 (m, 2H),0.77-0.79 (m, 2H).

Preparation of compound 70K (potassium salt of compound 70) potassium8-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(3-fluoro-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (84.0 mg, 0.23 mmol) to afford the K salt of compound 70 as ayellow solid (87.8 mg, 92%).

ESI-MS m/z: 354 (M−K+H)⁺, 352 (M−K−H)⁻.

Preparation of Compound 71 Preparation of tert-butyl5-bromo-3-cyclopropyl-1H-indazole-1-carboxylate

tert-Butyl 5-bromo-3-cyclopropyl-1H-indazole-1-carboxylate was preparedaccording to General Procedure F from 5-bromo-3-cyclopropyl-1H-indazole(500 mg, 2.11 mmol). tert-Butyl5-bromo-3-cyclopropyl-1H-indazole-1-carboxylate was obtained as acolorless oil (760 mg, 89%).

ESI-MS m/z: isotopic 339 and 337 (M+H)⁺.

Preparation of tert-butyl3-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-1-carboxylate

tert-Butyl3-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-1-carboxylatewas prepared according to General Procedure G from tert-butyl5-bromo-3-cyclopropyl-1H-indazole-1-carboxylate (760 mg, 1.87 mmol).Purification by flash silica column chromatography (heptane: ethylacetate) (1:0 to 3:1) afforded the title compound as a white solid (750mg, 100%).

ESI-MS m/z: 385 (M+H)⁺; 1H NMR (400 MHz, DMSO) δ ppm 8.23 (s, 1H), 8.04(d, J=8.6 Hz, 1H), 7.92 (dd, J=8.6 Hz, J=1.0 Hz, 1H), 2.23-2.31 (m, 1H),1.71 (s, 9H), 1.38 (s, 12H), 1.21-1.28 (m, 2H), 1.04-1.10 (m, 2H).

Preparation of methyl8-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (75mg, 0.26 mmol) and tert-butyl3-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-1-carboxylate(157.9 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as ayellow solid (84 mg, 60%).

ESI-MS m/z: 414 (M+H)⁺, 412 (M−H)⁻.

Preparation of compound 718-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(84 mg, 0.20 mmol) to afford compound 71 as a yellow solid (57 mg, 80%).

ESI-MS m/z: 400 (M+H)⁺, 398 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm12.87 (s, 1H), 9.35 (d, J=7.3 Hz, 1H), 8.25 (s, 1H), 8.01 (s, 1H), 7.70(d, J=7.3 Hz, 1H), 7.63 (d, J=8.6 Hz, 1H), 7.51 (dd, J=8.6 Hz, J=1.4 Hz,1H), 2.91 (s, 3H), 2.53-2.60 (m, 1H), 2.35-2.42 (m, 1H), 1.05-1.12 (m,3H), 0.97-1.04 (m, 4H), 0.79-0.84 (m, 2H).

Preparation of compound 71K (potassium salt of compound 71) potassium8-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(3-cyclopropyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (54.8 mg, 0.14 mmol) to afford the K salt of compound 71 as ayellow solid (54.3 mg, 89%).

ESI-MS m/z: 400 (M−K+H)⁺.

Preparation of Compound 72 Preparation of tert-Butyl5-bromo-2-hydroxybenzylcarbamate

tert-Butyl 5-bromo-2-hydroxybenzylcarbamate was prepared according toGeneral Procedure E from 5-bromo-2-hydroxybenzonitrile (1 g, 5.05 mmol).Purification by flash silica column chromatography (heptane: ethylacetate) (1:0 to 4:1) afforded the title compound as a white solid (376mg, 19%).

ESI-MS m/z: isotopic 302 and 300 (M+H)⁺.

Preparation of tert-butyl2-hydroxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamate

tert-Butyl2-hydroxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamatewas prepared according to General Procedure G from tert-Butyl5-bromo-2-hydroxybenzylcarbamate (376 mg, 1.24 mmol). Purification byflash silica column chromatography (heptane: ethyl acetate) (1:0 to 9:1)afforded the title compound as a white solid (96 mg, 21%).

SI-MS m/z: 348 (M−H)⁻; 1H NMR (400 MHz, DMSO) δ ppm 9.30 (s, 1H), 7.67(dd, J=8.1 Hz, J=1.3 Hz, 1H), 7.53 (d, J=1.5 Hz, 1H), 6.94 (d, J=8.1 Hz,1H), 5.23-5.26 (m, 1H), 4.23 (d, J=6.8 Hz, 2H), 1.43 (s, 9H), 1.33 (s,12H).

Preparation of methyl8-(3-(((tert-butoxycarbonyl)amino)methyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-(((tert-butoxycarbonyl)amino)methyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (65mg, 0.22 mmol) and tert-butyl2-hydroxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamate(93.3 mg, 0.27 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(85 mg, 100%).

ESI-MS m/z: 479 (M+H)⁺, 477 (M−H)⁻.

Preparation of8-(3-((tert-butoxycarbonylamino)methyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-((tert-Butoxycarbonylamino)methyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-(((tert-butoxycarbonyl)amino)methyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(85 mg, 0.22 mmol) to afford the title compound as a yellow solid (27mg, 27%).

ESI-MS m/z: 465 (M+H)⁺, 463 (M−H)⁻.

Preparation of compound 72HCl (hydrochloric salt of compound 72)8-(3-(aminomethyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

8-(3-(Aminomethyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride was prepared according to General Procedure C from8-(3-((tert-butoxycarbonylamino)methyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (27.0 mg, 0.06 mmol) to afford the HCl salt of compound 72 as ayellow solid (23.4 mg, 97%).

ESI-MS m/z: 365 (M+H)⁺, 363 (M−H)⁻; 1H NMR (400 MHz, MeOD-d6) δ ppm 9.40(s, 1H), 8.42 (s, 1H), 7.40-7.65 (m, 3H), 7.11 (d, J=7.8 Hz, 1H), 4.22(s, 2H), 2.96 (s, 3H), 2.45-2.60 (m, 1H), 1.05-1.20 (m, 2H), 0.75-0.90(m, 2H).

Preparation of compound 72K (potassium salt of compound 72) potassium8-(3-(aminomethyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-(aminomethyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(3-(aminomethyl)-4-hydroxyphenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (23.4 mg, 0.06 mmol) to afford the K salt of compound72 as a yellow solid (21.2 mg, 88%).

ESI-MS m/z: 365 (M−K+H)⁺, 363 (M−K−H)⁻.

Preparation of Compound 73 Preparation of a mixture of tert-butyl(5-bromobenzo[d]thiazol-2-yl)carbamate and di-tert-butyl(5-bromobenzo[d]thiazol-2-yl)di-carbamate

tert-Butyl (5-bromobenzo[d]thiazol-2-yl)carbamate and di-tert-butyl(5-bromobenzo[d]thiazol-2-yl)di-carbamate were prepared according toGeneral Procedure F from 5-bromobenzo[d]thiazol-2-amine (500 mg, 2.18mmol). The reaction yielded a 1:1 mixture of tert-butyl(5-bromobenzo[d]thiazol-2-yl)carbamate and di-tert-butyl(5-bromobenzo[d]thiazol-2-yl)di-carbamate as an off white solid (782 mg,83%). The mixture was not separated.

ESI-MS m/z: isotopic 331 and 329 (M+H)⁺ for mono-BOC component andisotopic 431 and 429 (M+H)⁺ for di-BOC component.

Preparation of a mixture of tert-butyl(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl)carbamateand di-tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl-di-carbamate

The titles compounds were made according to General Procedure G from amixture of tert-butyl (5-bromobenzo[d]thiazol-2-yl)carbamate anddi-tert-butyl (5-bromobenzo[d]thiazol-2-yl)di-carbamate (782 mg, 1.82mmol). Purification by flash silica column chromatography (heptane:ethyl acetate) (1:0 to 3:1) afforded a 1:1 mixture of tert-butyl(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl)carbamateand di-tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl-di-carbamateas white solid (774 mg, 97%).

ESI-MS m/z: 377 (M+H)⁺, 477 377 (M+H)⁺.

Preparation of methyl8-(2-(tert-butoxycarbonyl)amino)benzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylateand methyl8-(2-aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

The title compounds were prepared according to General Procedure A′ frommethyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and a mixture of tert-butyl(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl)carbamateand di-tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl-di-carbamate(195.7 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded methyl8-(2-aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(58 mg, 38%) and yellow solid methyl8-(2-(tert-butoxycarbonylamino)benzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(42 mg, 20%).

ESI-MS m/z: 406 (M+H)⁺ and 404 (M−H)⁻ for deprotected compound and 506(M+H)⁺ and 504 (M−H)⁻ for the BOC-protected compound.

Preparation of compound 738-(2-aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(2-Aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(2-amino)benzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(58 mg, 0.13 mmol) to afford compound 73 as a yellow solid (25.7 mg,55%).

ESI-MS m/z: 392 (M+H)⁺, 390 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.34(d, J=7.3 Hz, 1H), 8.87 (s, 2H), 8.27 (s, 1H), 7.98 (d, J=8.3 Hz, 1H),7.60 (d, J=7.3 Hz, 1H), 7.58 (d, J=1.3 Hz, 1H), 7.32 (dd, J=8.1 Hz,J=1.5 Hz, 1H), 2.88 (s, 3H), 2.52-2.58 (m, 1H), 1.05-1.10 (m, 2H),0.78-0.82 (m, 2H)

Preparation of compound 73K (potassium salt of compound 73) potassium8-(2-aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(2-aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(2-aminobenzo[d]thiazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (23.8 mg, 0.06 mmol) to afford the K salt of compound 73 as ayellow solid (21.6 mg, 80%).

ESI-MS m/z: 392 (M−K+H)⁺, 390 (M−K−H)⁻.

Preparation of Compound 74 Preparation of a mixture of tert-butyl5-bromo-1H-benzo[d]imidazole-1-carboxylate and tert-butyl6-bromo-1H-benzo[d]imidazole-1-carboxylate

The title compounds were made according to General Procedure F from5-bromo-1H-benzo[d]imidazole (500 mg, 2.54 mmol). Purification by flashsilica column chromatography (heptane: ethyl acetate) (1:0 to 1:1)afforded a mixture of tert-butyl5-bromo-1H-benzo[d]imidazole-1-carboxylate and tert-butyl6-bromo-1H-benzo[d]imidazole-1-carboxylate as a colorless oil (537 mg,70%).

ESI-MS m/z: isotopic 299 and 297 (M+H)⁺.

Preparation of a mixture of tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole-1-carboxylateand tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole-1-carboxylate

The title compounds were made according to General Procedure G from amixture of tert-butyl 5-bromo-1H-benzo[d]imidazole-1-carboxylate andtert-butyl 6-bromo-1H-benzo[d]imidazole-1-carboxylate (537 mg, 1.81mmol). Purification by flash silica column chromatography (heptane:ethyl acetate) (1:0 to 3:1) afforded a mixture of tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole-1-carboxylateand tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole-1-carboxylateas an orange oil: (937 mg, 98%).

ESI-MS m/z: 345 (M+H)⁺.

Preparation of methyl8-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (75mg, 0.26 mmol) and a mixture of tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole-1-carboxylateand tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole-1-carboxylate(141.5 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the titleBOC-deprotected compound as a yellow solid (69.9 mg, 65%).

ESI-MS m/z: 374 (M+H)⁺, 372 (M−H)⁻.

Preparation of compound 748-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(69.9 mg, 0.17 mmol) to afford compound 74 as a yellow solid (57.5 mg,94%).

ESI-MS m/z: 360 (M+H)⁺, 358 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.14 (s, 1H), 9.36 (d, J=7.3 Hz, 1H), 8.76 (s, 1H), 8.27 (s, 1H),7.85-7.87 (m, 2H), 7.67 (d, J=7.4 Hz, 1H), 7.46-7.49 (m, 1H), 2.89 (s,3H), 2.50-2.60 (m, 1H), 1.06-1.11 (m, 2H), 0.79-0.83 (m, 2H).

Preparation of compound 74K (potassium salt of compound 74) potassium8-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-benzo[d]imidazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (54.5 mg, 0.15 mmol) to afford the K salt of compound 74 as ayellow solid (47.3 mg, 76%).

ESI-MS m/z: 360 (M−K+H)⁺, 358 (M−K−H)⁻.

Preparation of Compound 75 Preparation of ethyl8-(1H-indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Ethyl8-(1H-indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from ethyl8-chloro-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(50 mg, 0.15 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (45.2 mg,0.19 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(48 mg, 77%).

ESI-MS m/z: 406 (M+H)⁺.

Preparation of compound 758-(1H-indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from ethyl1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (48mg, 0.12 mmol) to afford compound 75 as a yellow solid (36 mg, 78%).

ESI-MS m/z: 378 (M+H)⁺, 376 (M−H)⁻; 1H NMR (400 MHz, DMSO) δ ppm 13.37(s, 1H), 9.40 (d, J=5.6 Hz, 1H), 8.27 (s, 1H), 8.23 (s, 1H), 7.94 (s,1H), 5.96 (d, J=8.6 Hz, 1H), 3.83 (d, J=8.6 Hz, 1H), 2.83 (s, 3H),2.50-2.59 (m, 1H), 1.05-1.11 (m, 2H), 0.79-0.83 (m, 2H).

Preparation of compound 75K (potassium salt of compound 75) potassium8-(1H-indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylateaccording to General Procedure D from8-(1H-indazol-5-yl)-1-cyclopropyl-7-fluoro-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (30.0 mg, 0.08 mmol) to afford the K salt of compound 75 as ayellow solid (30.7 mg, 89%).

ESI-MS m/z: 354 (M−K+H)⁺, 352 (M−K−H)⁻.

Preparation of Compound 76 Preparation of tert-butylN-[4-bromo-2-[(tert-butoxycarbonylamino)-methyl]-phenyl]carbamate

tert-ButylN-[4-bromo-2-[(tert-butoxycarbonylamino)-methyl]-phenyl]-carbamate wasprepared according to General Procedure E from2-amino-5-bromo-benzonitrile (1.045 g, 5.30 mmol). Purification by flashsilica column chromatography (heptane: ethyl acetate) (1:0 to 9:1)afforded the title compound as a colorless oil (1.834 g, 84%).

ESI-MS m/z: isotopic 401 and 399 (M+H)⁺; 1H NMR (400 MHz, DMSO) δ ppm8.21 (br s, 1H), 7.95 (d, J=7.8 Hz, 1H), 7.37 (dd, J=8.8 Hz, J=2.3 Hz,1H), 7.26 (d, J=2.3 Hz, 1H), 5.00 (br s, 1H), 4.22 (d, J=6.6 Hz, 1H),1.53 (s, 9H), 1.46 (s, 9H).

Preparation of tert-butylN-[2-[(tert-butoxycarbonylamino)methyl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]carbamate

tert-ButylN-[2-[(tert-butoxycarbonylamino)methyl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-carbamatewas prepared according to General Procedure G from tert-butylN-[4-bromo-2-[(tert-butoxycarbonylamino)-methyl]-phenyl]-carbamate (780mg, 1.94 mmol). Purification by flash silica column chromatography(heptane: ethyl acetate) (1:0 to 9:1) afforded the title compound as awhite solid (314 mg, 30%).

ESI-MS m/z: 449 (M+H)⁺, 447 (M−H)⁻; 1H NMR (400 MHz, DMSO) δ ppm 8.39(br s, 1H), 8.16 (d, J=8.1 Hz, 1H), 7.72 (dd, J=8.3 Hz, J=1.2 Hz, 1H),7.55 (s, 1H), 4.85-5.00 (m, 1H), 4.28 (d, J=6.3 Hz, 2H), 1.52 (s, 9H),1.46 (s, 9H), 1.33 (s, 12H).

Preparation of methyl8-(4-(tert-butoxycarbonyl)amino)-3-((tert-butoxycarbonyl)amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(4-(tert-butoxycarbonyl)amino)-3-((tert-butoxycarbonyl)amino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (95mg, 0.33 mmol) and tert-butylN-[2-[(tert-butoxycarbonylamino)methyl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-carbamate(175 mg, 0.39 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(176 mg, 94%).

ESI-MS m/z: 578 (M+H)⁺.

Preparation of8-(4-(tert-butoxycarbonylamino)-3-((tert-butoxycarbonylamino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(4-(tert-Butoxycarbonylamino)-3-((tert-butoxycarbonylamino)-methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(4-((tert-butoxycarbonyl)amino)-3-((tert-butoxycarbonyl)-amino)methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(176 mg, 0.30 mmol) to afford the title compound as a yellow solid (123mg, 84%).

ESI-MS m/z: 564 (M+H)⁺, 562 (M−H)⁻.

Preparation of compound 76HCl (hydrochloric salt of compound 76)8-(4-amino-3-(aminomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

8-(4-Amino-3-(aminomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride was prepared according to General Procedure C from8-(4-(tert-butoxycarbonylamino)-3-((tert-butoxycarbonylamino)methyl)-phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (123.0 mg, 0.22 mmol) to afford the HCl salt of compound 76 as ayellow solid (54.7 mg, 60%).

ESI-MS m/z: 364 (M+H)⁺, 362 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.31(d, J=7.3 Hz, 1H), 8.31 (s, 3H), 8.22 (s, 1H), 7.60 (d, J=7.6 Hz, 1H),7.53 (d, J=1.8 Hz, 1H), 7.41 (dd, J=8.3 Hz, J=1.8 Hz, 1H), 6.95 (d,J=8.3 Hz, 1H), 4.04 (s, 2H), 2.92 (s, 3H), 2.40-2.57 (m, 1H), 1.06-1.11(m, 2H), 0.75-0.78 (m, 2H).

Preparation of compound 76K (potassium salt of compound 76) potassium8-(4-amino-3-(aminomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(4-amino-3-(aminomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(4-amino-3-(aminomethyl)phenyl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (51.7 mg, 0.13 mmol) to afford the K salt of compound76 as a yellow solid (47.9 mg, 86%).

ESI-MS m/z: 364 (M−K+H)⁺, 362 (M−K−H)⁻.

Preparation of Compound 77 Preparation of tert-butyl5-bromoindoline-1-carboxylate

tert-Butyl 5-bromoindoline-1-carboxylate was prepared according toGeneral Procedure F using 5-bromoindoline (500 mg, 2.52 mmol) and wasobtained as a brown solid; (769 mg, 63%).

ESI-MS m/z: isotopic 244 and 242 (M−tBu+H)⁺.

Preparation of tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline-1-carboxylate

tert-Butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline-1-carboxylatewas prepared according to General Procedure G from tert-butyl5-bromoindoline-1-carboxylate (769 mg, 1.60 mmol). Purification by flashsilica column chromatography (heptane: ethyl acetate) (1:0 to 3:1)afforded the title compound as a white solid (581 mg, 100%).

ESI-MS m/z: 290 (M−tBu+H)⁺; 1H NMR (400 MHz, DMSO) δ ppm 7.84 (br s,1H), 7.64 (d, J=7.8 Hz, 1H), 7.59 (s 1H), 3.97 (t, J=8.4 Hz, 2H), 3.07(t, J=8.7 Hz, 2H), 1.56 (s, 9H), 1.33 (s, 12H).

Preparation of methyl8-(1-(tert-butoxycarbonyl)indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1-(tert-butoxycarbonyl)indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (75mg, 0.26 mmol) and tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline-1-carboxylate(106.4 mg, 0.31 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as ayellow solid (121 mg, 99%).

ESI-MS m/z: 475 (M+H)⁺.

Preparation of8-(1-(tert-butoxycarbonyl)indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1-(tert-Butoxycarbonyl)indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1-(tert-butoxycarbonyl)indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(121 mg, 0.26 mmol) to afford compound 77 as a yellow solid (102.2 mg,89%).

ESI-MS m/z: 461 (M+H)⁺, 362 (M−H)⁻.

Preparation of compound 77HCl (hydrochloric salt of compound 77)8-(indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

8-(Indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride was prepared according to General Procedure C from8-(1-(tert-butoxycarbonyl)indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (102.2 mg, 0.22 mmol) to afford the HCl salt of compound 77 as ayellow solid (84.9 mg, 94%).

ESI-MS m/z: 361 (M+H)⁺, 359 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.30(d, J=7.6 Hz, 1H), 8.24 (s, 1H), 7.57 (d, J=7.3 Hz, 1H), 7.49 (s, 1H),7.38 (d, J=7.8 Hz, 1H), 7.16 (d, J=7.6 Hz, 1H), 3.69 (t, J=8.2 Hz, 2H),3.18 (t, J=8.2 Hz, 2H), 2.88 (s, 3H), 2.40-2.57 (m, 1H), 1.05-1.09 (m,2H), 0.76-0.79 (m, 2H).

Preparation of compound 77K (potassium salt of compound 77) potassium8-(indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(indolin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (82.9 mg, 0.21 mmol) to afford the K salt of compound77 as a yellow solid (99.9 mg, 100%).

ESI-MS m/z: 361 (M−K+H)⁺, 359 (M−K−H)⁻.

Preparation of Compound 78 Preparation of methyl8-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) andN-methyl-5-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)pyridin-2-amine (96.2mg, 0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(131.6 mg, 96%).

ESI-MS m/z: 364 (M+H)⁺.

Preparation of compound 788-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-(Methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(131.6 mg, 0.33 mmol) to afford compound 78 as a yellow solid (39.5 mg,31%).

ESI-MS m/z: 350 (M+H)⁺, 348 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.01 (s, 1H), 9.33 (d, J=7.3 Hz, 1H), 8.85 (s, 1H), 8.26 (s, 1H), 8.20(s, 1H), 8.01 (d, J=9.3 Hz, 1H), 7.61 (d, J=7.3 Hz, 1H), 7.11 (d, J=9.1Hz, 1H), 3.01 (s, 3H), 2.89 (s, 3H), 2.50-2.58 (m, 1H), 1.06-1.10 (m,2H), 0.78-0.82 (m, 2H).

Preparation of compound 78K (potassium salt of compound 78) potassium8-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-(methylamino)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (38.2 mg, 0.11 mmol) to afford the K salt of compound 78 as ayellow solid (37.9 mg, 89%).

ESI-MS m/z: 350 (M−K+H)⁺, 348 (M−K−H)⁻.

Preparation of Compound 79 Preparation of methyl8-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine(96.2 mg, 0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded quantitatively the title compound as ayellow solid.

ESI-MS m/z: 364 (M+H)⁺.

Preparation of compound 798-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-Amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.34 mmol) to afford compound 79 as a yellow solid (75 mg, 57% in twosteps).

ESI-MS m/z: 350 (M+H)⁺, 348 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.13 (s, 1H), 9.28 (d, J=7.0 Hz, 1H), 8.21 (s, 1H), 8.07 (s, 1H), 7.60(d, J=6.8 Hz, 1H), 7.54 (s, 1H), 6.33 (s, 2H), 2.91 (s, 3H), 2.40-2.60(m, 1H), 2.14 (s, 3H), 1.06-1.09 (m, 2H), 0.72-0.80 (m, 2H).

Preparation of compound 79K (potassium salt of compound 79) potassium8-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-amino-5-methylpyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (71.2 mg, 0.20 mmol) to afford the K salt of compound 79 as ayellow solid (81.1 mg, 100%).

ESI-MS m/z: 350 (M−K+H)⁺.

Preparation of Compound 80 Preparation of methyl8-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (3-methyl-1H-indazol-5-yl)boronic acid (176.1 mg,0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(120 mg, 88%).

ESI-MS m/z: 388 (M+H)⁺.

Preparation of compound 808-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(120 mg, 0.30 mmol) to afford compound 80 as a yellow solid (97 mg,84%).

ESI-MS m/z: 374 (M+H)⁺, 372 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.35(d, J=7.3 Hz, 1H), 8.25 (s, 1H), 7.96 (s, 1H), 7.68 (d, J=7.3 Hz, 1H),7.64 (d, J=8.6 Hz, 1H), 7.51 (dd, J=1.5 Hz, J=8.6 Hz, 1H), 2.90 (s, 3H),2.50-2.60 (m, 1H), 2.56 (s, 3H), 1.06-1.10 (m, 2H), 0.79-0.83 (m, 2H).

Preparation of compound 80K (potassium salt of compound 80) potassium8-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(3-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (92.2 mg, 0.25 mmol) to afford the K salt of compound 80 as ayellow solid (103.3 mg, 100%).

ESI-MS m/z: 374 (M+H)⁺, 372 (M−H)⁻.

Preparation of Compound 81 Preparation of methyl8-(1-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and (1-methyl-1H-indazol-5-yl)boronic acid (176.0 mg,0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(129.8 mg, 95%).

ESI-MS m/z: 388 (M+H)⁺.

Preparation of compound 818-(1-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1-Methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl1-cyclopropyl-9-methyl-8-(1-methyl-1H-indazol-5-yl)-4-oxo-4H-quinolizine-3-carboxylate(129.8 mg, 0.33 mmol) to afford compound 81 as a yellow solid (104.2 mg,77%).

ESI-MS m/z: 374 (M+H)⁺, 372 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.12 (s, 1H), 9.34 (d, J=7.3 Hz, 1H), 8.26 (s, 1H), 8.20 (s, 1H), 8.00(s, 1H), 7.85 (d, J=8.6 Hz, 1H), 7.66 (d, J=7.1 Hz, 1H), 7.59 (d, J=8.4Hz, 1H), 4.13 (s, 3H), 2.89 (s, 3H), 2.50-2.60 (m, 1H), 1.06-1.10 (m,2H), 0.79-0.81 (m, 2H).

Preparation of compound 81K (potassium sal t of compound 81) potassium8-(1-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1-methyl-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from1-cyclopropyl-9-methyl-8-(1-methyl-1H-indazol-5-yl)-4-oxo-4H-quinolizine-3-carboxylicacid (97.2 mg, 0.26 mmol) to afford the K salt of compound 81 as ayellow solid (101.5.0 mg, 87%).

ESI-MS m/z: 374 (M−K+H)⁺, 372 (M−K−H)⁻.

Preparation of Compound 82 Preparation of methyl8-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 1H-Indazole-4-boronic acid (67 mg, 0.41 mmol).Purification by flash silica column chromatography (DCM:MeOH) (1:0 to9:1) afforded the title compound as a yellow solid (110.5 mg, 79%).

ESI-MS m/z: 374 (M+H)⁺.

Preparation of compound 828-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(110.5 mg, 0.27 mmol) to afford compound 82 as a yellow solid (85.2 mg,73%).

ESI-MS m/z: 360 (M+H)⁺, 358 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.13 (s, 1H), 13.44 (s, 1H), 9.40 (d, J=7.4 Hz, 1H), 8.30 (s, 1H), 7.95(s, 1H), 7.73 (d, J=8.3 Hz, 1H), 7.65 (d, J=7.4 Hz, 1H), 7.56 (t, J=7.7Hz, 1H), 7.25 (d, J=6.8 Hz, 1H), 2.78 (s, 3H), 2.54-2.61 (m, 1H),1.04-1.10 (m, 2H), 0.80-0.87 (m, 2H).

Preparation of compound 82K (potassium salt of compound 82) potassium8-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-indazol-4-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (30.0 mg, 0.08 mmol) to afford the K salt of compound 82 as ayellow solid (30.5 mg, 92%).

ESI-MS m/z: 360 (M−K+H)⁺, 358 (M−K−H)⁻.

Preparation of Compound 83 Preparation of methyl8-(1H-indazol-6-yl)-91-cyclopropyl-1-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-indazol-6-yl)-91-cyclopropyl-1-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and 1H-indazole-6-boronic acid pinacol ester (103 mg,0.41 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(121.2 mg, 86%).

ESI-MS m/z: 374 (M+H)⁺.

Preparation of compound 838-(1H-indazol-6-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indazol-6-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-indazol-6-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(121.2 mg, 0.30 mmol) to afford compound 83 as a yellow solid (51.7 mg,43%).

ESI-MS m/z: 360 (M+H)⁺, 358 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm14.11 (s, 1H), 13.35 (s, 1H), 9.36 (d, J=7.3 Hz, 1H), 8.28 (s, 1H), 8.21(s, 1H), 7.96 (d, J=8.2 Hz, 1H), 7.70 (s, 1H), 7.66 (d, J=7.3 Hz, 1H),7.26 (d, J=8.2 Hz, 1H), 2.88 (s, 3H), 2.50-2.59 (m, 1H), 1.05-1.10 (m,2H), 0.79-0.83 (m, 2H).

Preparation of compound 83K (potassium salt of compound 83) potassium8-(1H-indazol-6-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-indazol-6-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-indazol-6-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (45.1 mg, 0.13 mmol) to afford the K salt of compound 83 as ayellow solid (49.6 mg, 97%).

ESI-MS m/z: 360 (M+H)⁺, 358 (M−H)⁻.

Preparation of Compound 84 Preparation of methyl8-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and tert-butyl4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperazine-1-carboxylate(159.9 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded quantitatively the titlecompound as a yellow solid.

ESI-MS m/z: 519 (M+H)⁺.

Preparation of8-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-(4-(tert-Butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(0.34 mmol) to afford the title compound 84 as a yellow solid (144.6 mg,100%).

ESI-MS m/z: 505 (M+H)⁺.

Preparation of compound 84HCl (hydrochloric salt of compound 84)8-(6-(piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride

8-(6-(Piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride was prepared according to General Procedure C from8-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (144.6 mg, 0.29 mmol) to afford the hydrochloric salt of compound84 as a yellow solid (122.7 mg, 97%).

ESI-MS m/z: 405 (M+H)⁺, 403 (M−H)⁻; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.40(s, 2H), 9.30 (d, J=7.3 Hz, 1H), 8.40 (d, J=2.2 Hz, 1H), 8.23 (s, 1H),7.92 (d, J=9.1 Hz, J=2.5 Hz, 1H), 7.63 (d, J=7.6 Hz, 1H), 7.15 (d, J=8.8Hz, 1H), 3.89-3.92 (m, 4H), 3.18-3.26 (m, 4H), 2.90 (s, 3H), 2.50-2.57(m, 1H), 1.05-1.11 (m, 2H), 0.76-0.81 (m, 2H).

Preparation of compound 84K (potassium salt of compound 84) potassium8-(6-(piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-(piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-(piperazin-1-yl)pyridin-3-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid hydrochloride (122.7 mg, 0.28 mmol) to afford the K salt ofcompound 84 as a yellow solid (137.1 mg, 100%).

ESI-MS m/z: 405 (M−K+H)⁺, 403 (M−K−H)⁻.

Preparation of Compound 85 Preparation of methyl8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A′ from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine(100.3 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as ayellow solid (124.8 mg, 90%).

ESI-MS m/z: 374 (M+H)⁺.

Preparation of compound 858-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(124.8 mg, 0.31 mmol) to afford compound 85 as a yellow solid (26 mg,21%).

ESI-MS m/z: 360 (M+H)⁺, 358 (M−H)⁻; 1H NMR (400 MHz, MeOD-d6) δ ppm 9.43(d, J=7.3 Hz, 1H), 8.64 (s, 1H), 8.58 (s, 1H), 8.43 (s, 1H), 7.73 (d,J=3.5 Hz, 1H), 7.58 (d, J=7.6 Hz, 1H), 6.88 (d, J=3.5 Hz, 1H), 2.98 (s,3H), 2.50-2.58 (m, 1H), 1.13-1.18 (m, 2H), 0.86-0.90 (m, 2H).

Preparation of compound 85K (potassium salt of compound 85) potassium8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid (26.0 mg, 0.07 mmol) to afford the K salt of compound 85 as ayellow solid (21.1 mg, 70%).

ESI-MS m/z: 360 (M−K+H)⁺.

Preparation of Compound 86 Preparation of5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-amine

Compound5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-amine wasprepared according to General Procedure G from5-bromo-1H-indazol-3-amine (500 mg, 2.36 mmol). Purification by flashsilica column chromatography (DCM: MeOH) (1:0 to 94:6) andrecrystallization in DCM and heptane gave the title compound as a brownsolid (230 mg, 25%).

ESI-MS m/z: 260 (M+H)⁺.

Preparation of methyl8-(3-amino-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(3-amino-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate (100mg, 0.34 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-amine(106.4 mg, 0.41 mmol). Purification by flash silica columnchromatography (DCM:MeOH) (1:0 to 9:1) afforded the title compound as ayellow solid (32 mg, 42%).

ESI-MS m/z: 389 (M+H)⁺, 387 (M−H)⁻.

Preparation of Compound 868-(3-amino-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(3-Amino-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(3-amino-1H-indazol-5-yl)-1-cyclopropyl-9-methyl-4-oxo-4H-quinolizine-3-carboxylate(32 mg, 0.14 mmol). Purification by preparative HPLC afforded compound86 as a yellow solid (0.7 mg, 1%).

ESI-MS m/z: 375 (M+H)⁺; 1H NMR (400 MHz, CD₃OD) δ ppm 9.41 (d, J=7.3 Hz,1H), 8.42 (s, 1H), 7.92 (s, 1H), 7.43-7.52 (m, 3H), 2.97 (s, 3H),2.49-2.58 (m, 1H), 1.09-1.15 (m, 2H), 0.81-0.86 (m, 2H).

Preparation of Compound 87 Preparation of methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl 8-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate was prepared according toGeneral Procedure A from methyl8-chloro-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate(50 mg, 0.16 mmol) and 2-amino-pyridine-5-boronic acid pinacol ester (43mg, 0.20 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 25:4) afforded the title compound as a yellow solid(44 mg, 69%).

ESI-MS m/z: 3864 (M+H)⁺.

Preparation of compound 878-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-Amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from8-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate(44 mg, 0.12 mmol) to afford compound 87 as a yellow solid (19 mg, 41%).

ESI-MS m/z: 350 (M+H)⁺; 1H NMR (400 MHz DMSO-d6) δ ppm 14.2 (br s, 1H),9.26 (s, 1H), 8.19 (s, 1H), 7.95 (s, 1H), 7.45-7.67 (m, 2H), 7.20 (br s,2H), 6.86 (d, J=8.4 Hz, 1H), 2.73 (s, 3H), 2.50-2.60 (m, 1H), 2.20 (s,3H), 0.98-1.12 (m, 2H), 0.71-0.79 (m, 2H).

Preparation of compound 87K (potassium salt of compound 87) potassium8-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-amino-pyridin-3-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylicacid (29 mg, 0.08 mmol) to afford the K salt of compound 87 as a yellowsolid (28 mg, 90%).

ESI-MS m/z: 350 (M−K+H)⁺.

Preparation of Compound 88 Preparation of methyl8-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate(50 mg, 0.17 mmol), and 1H-Indazole-5-boronic acid pinacol ester (48 mg,0.19 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as a yellow solid(30 mg, 45%).

ESI-MS m/z: 388 (M+H)⁺.

Preparation of compound 888-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate(30 mg, 0.08 mmol) to afford compound 88 as a yellow solid (22 mg, 77%).

ESI-MS m/z: 374 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 14.25 (s, 1H),10.20 (br s, 1H), 9.32 (s, 1H), 8.54 (s, 1H), 8.19 (s, 1H), 7.71 (d,J=8.6 Hz, 1H), 7.59 (s, 1H), 7.26 (s, 1H), 7.18 (d, J=8.5 Hz, 1H), 2.73(s, 3H), 2.33-2.43 (m, 1H), 2.14 (s, 3H), 1.02-1.10 (m, 2H), 0.82-0.87(m, 2H).

Preparation of compound 88K (potassium salt of compound 88) potassium8-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(1H-indazol-5-yl)-1-cyclopropyl-7,9-dimethyl-4-oxo-4H-quinolizine-3-carboxylicacid (20 mg, 0.05 mmol) to afford the K salt of compound 88 as a yellowsolid (19 mg, 82%).

ESI-MS m/z: 374 (M−K+H)⁺.

Preparation of Compound 89 Preparation of methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate (50mg, 0.16 mmol), and 2-aminopyridine-5-boronic acid pinacol ester (42.9mg, 0.20 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as an orange solid(35 mg, 57%).

ESI-MS m/z: 366 (M+H)⁺.

Preparation of compound 898-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylicacid

8-(6-Amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate(34 mg, 0.09 mmol) to afford compound 89 as an orange solid (22.3 mg,68%).

ESI-MS m/z: 352 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 14.09 (br s, 1H),9.29 (d, J=7.6 Hz, 1H), 8.51 (d, J=2 Hz, 1H), 8.42 (s, 1H), 7.94 (dd,J=8.8 Hz, J=2.5 Hz 1H), 7.38 (d, J=7.3 Hz, 1H), 6.67 (d, J=8.8 Hz, 1H),4.84 (br s, 2H), 3.58 (s, 3H), 2.62-2.70 (m, 1H), 1.01-1.06 (m, 2H),0.81-0.90 (m, 2H).

Preparation of compound 89K (potassium salt of compound 89) potassium8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from8-(6-amino-pyridin-3-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylicacid (21.6 mg, 0.06 mmol) to afford the K salt of compound 89 as ayellow solid (22.7 mg, 91%).

ESI-MS m/z: 352 (M−K+H)⁺.

Preparation of Compound 90 Preparation of methyl8-(1H-indazol-5-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate

Methyl8-(1H-indazol-5-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure A from methyl8-chloro-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate (50mg, 0.16 mmol), and 1H-Indazole-5-boronic acid pinacol ester (47.6 mg,0.20 mmol). Purification by flash silica column chromatography(DCM:MeOH) (1:0 to 9:1) afforded the title compound as an orange foam(32 mg, 46%).

ESI-MS m/z: 390 (M+H)⁺.

Preparation of compound 908-(1H-indazol-5-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylicacid

8-(1H-Indazol-5-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylicacid was prepared according to General Procedure B from methyl1-cyclopropyl-8-(1H-indazol-5-yl)-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate(32 mg, 0.08 mmol) to afford compound 90 as a yellow solid (22.5 mg,73%).

ESI-MS m/z: 376 (M+H)⁺; 1H NMR (400 MHz, CDCl₃) δ ppm 14.12 (br s, 1H),13.37 (s, 1H), 9.27 (d, J=7.6 Hz, 1H), 8.25 (s, 2H), 8.13 (s, 1H),7.73-7.83 (m, 3H), 3.46 (s, 3H), 2.63-2.67 (m, 1H), 0.98-1.03 (m, 2H),0.74-0.78 (m, 2H).

Preparation of compound 90K (potassium salt of compound 90) potassium8-(1H-indazol-5-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylate

Potassium8-(1H-indazol-5-yl)-1-cyclopropyl-9-methoxy-4-oxo-4H-quinolizine-3-carboxylatewas prepared according to General Procedure D from1-cyclopropyl-8-(1H-indazol-5-yl)-9-methoxy-4-oxo-4H-quinolizine-3-carboxylicacid (15.3 mg, 0.04 mmol) to afford the K salt of compound 90 as ayellow solid (14.3 mg, 77%).

ESI-MS m/z: 376 (M−K+H)⁺.

1-4. (canceled)
 5. A pharmaceutical composition comprising Polymyxin Band a 4-oxoquinolizine compound of formula IIIc:

or a pharmaceutically acceptable salt thereof, wherein R₁ is hydrogen,halogen, cyano, C₁₋₈ alkyl, C₁₋₈ haloalkyl, —OR^(X), —N(R^(X))₂,—C(O)R^(X), —C(O)OR^(X), or —C(O)N(R^(X))₂, wherein each Rx isindependently hydrogen, C₁₋₈ alkyl, or C₁₋₈haloalkyl; and Y is aryl orheteroaryl, each of which is substituted with either —N(R^(Y1))₂ or—C₁-C₈ alkyl-R^(Y) where R^(Y) is —N(R^(Y1))₂ and further optionallysubstituted by one to four groups that are each independently halogen,C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y),wherein R^(Y) is nitro, cyano, —OR^(Y1), —SR^(Y1), —N(R^(Y1))₂,—C(O)R^(Y1), —C(O)OR^(Y1), —C(O)N(R^(Y1))₂, —OC(O)R^(Y1), —OC(O)OR^(Y1),—OC(O)N(R^(Y1))₂, —N(R^(Y1))C(O)R^(Y1), —N(R^(Y1))C(O)OR^(Y1),—N(R^(Y1))C(O)N(R^(Y1))₂, —S(O)₂R^(Y1), —S(O)₂OR^(Y1), —S(O)₂N(R^(Y1))₂,—OS(O)₂R^(Y1), —OS(O)₂OR^(Y1), —OS(O)₂N(R^(Y1))₂, —N(R^(Y1))S(O)₂R^(Y1),—N(R^(Y1))S(O)₂OR^(Y1), or —N(R^(Y1))S(O)₂N(R^(Y1))₂, wherein eachR^(Y1) is independently hydrogen, C₁₋₈ is alkyl, or C₁₋₈ haloalkyl. 6-7.(canceled)
 8. The pharmaceutical composition according to claim 5,wherein R₁ is hydrogen or halogen.
 9. (canceled)
 10. The pharmaceuticalcomposition according to claim 5, wherein Y is phenyl substituted withone group which is halogen, cyano, —OR^(Y1), —SR^(Y1), —N(R^(Y1))₂, C₁₋₈alkyl, —C₁₋₈ alkyl-N(R^(Y1))₂, or —C₁₋₈ alkyl-OR^(Y1), and optionallysubstituted by one or two groups that are each independently halogen,C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y);should Y is a 5-membered or 6-membered heteroaryl optionally substitutedby one to five groups that are each independently halogen, C₁₋₈ alkyl,C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl, heteroaryl,C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl, aryl(C₁₋₈)alkyl,heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y); or Y is a bicyclicheteroaryl optionally substituted by one to five groups that are eachindependently halogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl,heterocyclyl, aryl, heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl,heterocyclyl(C₁₋₈)alkyl, aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y),or —C₁₋₈ alkyl-R^(Y). 11-69. (canceled)
 70. A pharmaceutical compositionaccording to claim 8, wherein Y is phenyl substituted with one groupwhich is halogen, cyano, —OR^(Y1), —SR^(Y1), —N(R^(Y1))₂, C₁₋₈ alkyl,—C₁₋₈ alkyl-N(R^(Y1))₂, or —C₁₋₈ alkyl-OR^(Y1), and optionallysubstituted by one or two groups that are each independently halogen,C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl,aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y);should Y is a 5-membered or 6-membered heteroaryl optionally substitutedby one to five groups that are each independently halogen, C₁₋₈ alkyl,C₁₋₈ haloalkyl, C₃₋₈cycloalkyl, heterocyclyl, aryl, heteroaryl,C₃₋₈cycloalkyl(C₁₋₈) alkyl, heterocyclyl(C₁₋₈)alkyl, aryl(C₁₋₈)alkyl,heteroaryl(C₁₋₈)alkyl, —R^(Y), or —C₁₋₈ alkyl-R^(Y); or Y is a bicyclicheteroaryl optionally substituted by one to five groups that are eachindependently halogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈cycloalkyl,heterocyclyl, aryl, heteroaryl, C₃₋₈cycloalkyl(C₁₋₈) alkyl,heterocyclyl(C₁₋₈)alkyl, aryl(C₁₋₈)alkyl, heteroaryl(C₁₋₈)alkyl, —R^(Y),or —C₁₋₈ alkyl-R^(Y).
 71. A pharmaceutical composition according toclaim 5, wherein the 4-oxoquinolizine compound is selected from thegroup consisting of: Compound 1

8-(3-fluoro-4-amino-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 4

8-(3-fluoro-4-aminomethyl- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 5

8-(4-aminomethyl-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 6

8-(4-aminomethyl-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 7

8-(4-amino-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 8

8-(4-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 9

8-(3-amino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 11

8-(2-chloro-4-amino-5- methyl-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 12

8-[5-aminomethyl)-2-furyl]- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 13

8-[5-aminomethyl)-2- thienyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 16

8-(4-amino-3-ethyl-5-methyl- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 17

8-(3-fluoro-4-amino-phenyl)- 1-cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 18

8-(3-amino-phenyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 20

8-(2-fluoro-4-amino-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 21

8-(3-amino-4-fluoro-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 22

8-(3-amino-5-fluoro-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 24

8-(3-chloro-4-amino- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 25

8-(3-methoxy-4-amino- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 28

8-(4-methylamino-phenyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 31

8-(3-methyl-4-amino- phenyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 33

8-(6-amino-3-pyridyl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 34

8-(1H-indol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 35

8-(1H-indazol-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 37

8-(4-dimethylamino-phenyl)- 1-cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 57

8-(2-aminopyrimidin-5-yl)-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 60

8-(6-amino-3-pyridyl)-1- cyclopropyl-7-fluoro-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 69

8-[3-methyl-4- (methylamino)phenyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 72

8-[3-(aminomethyl)-4- hydroxy-phenyl]-1- cyclopropyl-9-methyl-4-oxo-quinolizine-3-carboxylic acid Compound 76

8-[3-(aminomethyl)-4-amino- phenyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 78

8-[6-(methylamino)-3- pyridyl]-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 79

8-(6-amino-5-methyl-3- pyridyl)-1-cyclopropyl-9-methyl-4-oxo-quinolizine-3- carboxylic acid Compound 87

8-(6-amino-3-pyridyl)-1- cyclopropyl-7,9-dimethyl-4-oxo-quinolizine-3-carboxylic acid Compound 89

8-(6-amino-3-pyridyl)-1- cyclopropyl-9-methoxy-4-oxo-quinolizine-3-carboxylic acid


72. A pharmaceutical composition according to claim 5, wherein thePolymyxin B is present in a subinhibitory concentration.
 73. A methodfor treatment of a bacterial infection in an individual in need thereofcomprising administering to the individual a pharmaceutical compositionaccording claim
 5. 74. A method for treatment of a bacterial infectionin an individual in need thereof, wherein the bacterial infection isinfection by a multiresistant strain, comprising administering to theindividual a pharmaceutical composition according to claim
 5. 75. Amethod for treatment of a bacterial infection in an individual in needthereof, wherein the infection is infection by one or more bacteria of agenus selected from the group consisting of Acinetobacter, Bacillus,Bortadella, Borrelia, Brucella, Camphylobacter, Chlamydia, Clostridium,Corynebacterium, Enterococcus, Escherichia, Fransisella, Haemophilus,Helicobacter, Legionella, Leptospira, Listeria, Mycobacterium,Mycoplasma, Neisseria, Propionibacterium, Pseudomonas, Rickettsia,Salmonella, Shigella, Staphylococcus, Streptococcus, Treponema, Vibrioand Yersinia, comprising administering to the individual apharmaceutical composition according to claim 5.